Nutritional composition having lipophilic compounds with improved solubility and bioavailability

ABSTRACT

Disclosed herein is a nutritional composition having at least one protein, at least one fat, and at least one lipophilic compound, the composition comprising at least one assembly comprising at least one hydrophobic protein, monoglycerides and diglycerides (“MDG”) and at least one lipophilic compound, wherein at least 1% of the total MDG in the nutritional composition remains in the aqueous phase after centrifugation at 100,000×g for 1 hour at 20° C.

RELATED APPLICATION INFORMATION

This application claims the benefit of U.S. Application No. 61/871,719,filed on Aug. 29, 2013, U.S. Application No. 61/901,207, filed on Nov.7, 2013, U.S. Application No. 61/920,657, filed on Dec. 24, 2013, U.S.Application No. 61/920,666, filed on Dec. 24, 2013, U.S. Application No.61/920,669 filed on Dec. 24, 2013, U.S. Application No. 61/920,675,filed on Dec. 24, 2013, U.S. Application No. 61/949,505 filed on Mar. 7,2014 and U.S. Application No. 62/007,037 filed on Jun. 3, 2014, thecontents of each of which are herein incorporated by reference in itsentirety.

TECHNICAL FIELD

The present disclosure relates to compositions containing lipophiliccompounds with improved bioavailability. More particularly, thecompositions contain an activated premix or an MDG-protected premixcomprising monoglycerides and diglycerides. These premixes may providelipophilic compounds in a form that is more available for absorptioninto the body upon consumption.

BACKGROUND

Adult, pediatric, and infant manufactured nutritional and pharmaceuticalproducts may comprise a variety of lipophilic compounds such ascarotenoids, lipid soluble vitamins, lipophilic antioxidants orcombinations thereof. Current nutritional compositions comprisinglipophilic compounds typically provide limited bioavailability followingconsumption because lipophilic compounds are unable to remain in stable,water soluble forms. As a result, the products are often over-fortifiedwith lipophilic compounds to ensure that the desired nutritional orpharmaceutical benefits from the compounds may be obtained. In somecases, the over-fortification may be from about 2 times to about 10times the amount required to achieve the desired benefits.

These high fortification rates may lead to increased production costsand potential complications in formulating the final product withoutproviding additional consumer benefits. Furthermore, the over fortifiedformulations may overwhelm the digestive tract with non-solublelipophilic compounds leading to indigestion.

Accordingly, there is a need for nutritional compositions for infants,toddlers, children and adults that provide necessary lipophiliccompounds in a more stable soluble form.

SUMMARY OF THE INVENTION

In one aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein is3-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein is3-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof wherein the assembly has a size of 60 kD to 400kD.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w).

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof; wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w).

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein is3-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition,further comprising lecithin.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition,further comprising lecithin, comprising about 10 mg/kg to about 5 g/kgof lecithin based on total weight of the composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition,further comprising lecithin, comprising about 10 mg/kg to about 5 g/kgof lecithin based on total weight of the composition, further comprisingdocosahexaenoic acid (“DHA”).

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition,further comprising lecithin, comprising about 10 mg/kg to about 5 g/kgof lecithin based on total weight of the composition, further comprisingdocosahexaenoic acid (“DHA”), wherein the nutritional compositioncomprises about 10 mg/kg to about 10 g/kg DHA based on total weight ofthe composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition,further comprising lecithin, comprising about 10 mg/kg to about 5 g/kgof lecithin based on total weight of the composition, further comprisingdocosahexaenoic acid (“DHA”), wherein the nutritional compositioncomprises about 10 mg/kg to about 10 g/kg DHA based on total weight ofthe composition, further comprising high palmitic acid vegetable oil.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein is3-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition,further comprising lecithin, comprising about 10 mg/kg to about 5 g/kgof lecithin based on total weight of the composition, further comprisingdocosahexaenoic acid (“DHA”), wherein the nutritional compositioncomprises about 10 mg/kg to about 10 g/kg DHA based on total weight ofthe composition, further comprising high palmitic acid vegetable oil,wherein the nutritional composition comprises about 0.5 g/kg to about 35g/kg of high palmitic acid vegetable oil based on total weight of thecomposition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein is3-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition,further comprising lecithin, comprising about 10 mg/kg to about 5 g/kgof lecithin based on total weight of the composition, further comprisingdocosahexaenoic acid (“DHA”), wherein the nutritional compositioncomprises about 10 mg/kg to about 10 g/kg DHA based on total weight ofthe composition, further comprising high palmitic acid vegetable oil,wherein the nutritional composition comprises about 0.5 g/kg to about 35g/kg of high palmitic acid vegetable oil based on total weight of thecomposition, wherein the nutritional composition comprises about 0.2 g/Lto about 20 g/L of β-casein based on total weight of the composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein isβ-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition,further comprising lecithin, comprising about 10 mg/kg to about 5 g/kgof lecithin based on total weight of the composition, further comprisingdocosahexaenoic acid (“DHA”), wherein the nutritional compositioncomprises about 10 mg/kg to about 10 g/kg DHA based on total weight ofthe composition, further comprising high palmitic acid vegetable oil,wherein the nutritional composition comprises about 0.5 g/kg to about 35g/kg of high palmitic acid vegetable oil based on total weight of thecomposition, wherein the nutritional composition comprises about 0.2 g/Lto about 20 g/L of β-casein based on total weight of the composition,wherein the nutritional composition comprises a carbohydrate.

In another aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 1% of the total MDG in thenutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C., wherein the assemblyis (i) water soluble; and (ii) has a size of 100 kD to 1000 kD, whereinMDG increases the concentration of protein of the assembly by at least2% compared to a nutritional composition without MDG, wherein MDGincreases the bound lipophilic compound of the assembly by at least 2%compared to a nutritional composition without MDG, wherein thelipophilic compound of the composition has at least a 15% improvement inbioavailability when administered to a subject, wherein the 15%improvement in bioavailability is measured by lymphatic absorption ofthe composition over a nutritional composition without MDG, wherein theassembly is still water soluble after centrifugation at about 31,000×gfor at least 1 hour at 20° C., wherein at least 5% of the assemblyremains in the aqueous phase after centrifugation, wherein the proteinis intact protein, hydrolyzed protein or combinations thereof, whereinthe assembly is stable for at least 12 months at 23° C., wherein theassembly is stable for at least 24 months at room temperature, whereinthe assembly comprises a hydrophobic protein having a GRAVY value ofabout −0.5 to about 0, wherein the hydrophobic protein is3-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein, α-lactalbumin,or combinations thereof, wherein the assembly has a size of 60 kD to 400kD, wherein the lipophilic compound is a carotenoid, a lipid solublevitamin, a lipophilic antioxidant or combinations thereof, wherein thelipophilic compound is lutein, vitamin A, vitamin D, vitamin E, vitaminK or any combinations thereof, wherein the fat is MDG, coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic coconut oil, fractionated coconut oil, soy oil, corn oil,olive oil, safflower oil, high oleic safflower oil, sunflower oil, higholeic sunflower oil, palm oil, palm kernel oil, palm olein, canola oil,marine oils, cottonseed oil, flax seed oil, hemp seed oil, peanut oil,borage oil, algal oils, fungal oils, MCT oil (medium chaintriglycerides) and combinations thereof, comprising about 145 mg/kg toabout 890 mg/kg of MDG based on total weight of the composition,comprising about 145 mg/kg to about 600 mg/kg of MDG based on totalweight of the composition, comprising about 1.0 μg/kg to about 7.0 g/kgof a lipophilic compound based on total weight of the composition,comprising about 147.4 mg/kg to about 589.6 mg/kg of MDG and about 1.12mg/kg of a lipophilic compound based on total weight of the composition,wherein the ratio of MDG to lipophilic compound are selected from thegroup consisting of 12000:1, 11000:1, 10000:1, 9000:1, 8000:1, 7000:1,6000:1, 5000:1, 4000:1, 3000:1, 2000:1, 1000:1, 900:1, 800:1, 700:1,600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 75:1, 50:1, 25:1, 20:1, 15:1,14:1, 13:1, 12:1, 11:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 and1:1 (w/w), wherein the ratio of MDG to lipophilic compound comprises aratio of about 12000:1 to about 1:1 (w/w), wherein the ratio of MDG tolipophilic compound is 4:1, 3:1 or 2:1, comprising about 1.0 g/L toabout 100 g/L of hydrophobic protein based on total weight of thecomposition, further comprising choline, comprising about 5 mg/kg toabout 1 g/kg of choline based on total weight of the composition,further comprising lecithin, comprising about 10 mg/kg to about 5 g/kgof lecithin based on total weight of the composition, further comprisingdocosahexaenoic acid (“DHA”), wherein the nutritional compositioncomprises about 10 mg/kg to about 10 g/kg DHA based on total weight ofthe composition, further comprising high palmitic acid vegetable oil,wherein the nutritional composition comprises about 0.5 g/kg to about 35g/kg of high palmitic acid vegetable oil based on total weight of thecomposition, wherein the nutritional composition comprises about 0.2 g/Lto about 20 g/L of β-casein based on total weight of the composition,wherein the nutritional composition comprises a carbohydrate, whereinthe carbohydrate is maltodextrin, hydrolyzed or modified starch,hydrolyzed or modified cornstarch, glucose polymers, corn syrup, cornsyrup solids, rice-derived carbohydrates, glucose, fructose, lactose,high fructose corn syrup, honey, sugar alcohols, maltitol, erythritol,sorbitol, or combinations thereof.

In one aspect, the disclosure may be directed to a nutritionalcomposition comprising at least one protein and vitamin D, havingimproved bioavailability of vitamin D, wherein the nutritionalcomposition comprises: an assembly comprising a combination ofmonoglycerides and diglycerides (“MDG”) and vitamin D, wherein thevitamin D has improved bioavailability when administered to a subject.

In another aspect, the disclosure may be directed to a nutritionalcomposition comprising at least one protein and vitamin D, havingimproved bioavailability of vitamin D, wherein the nutritionalcomposition comprises: an assembly comprising a combination ofmonoglycerides and diglycerides (“MDG”) and vitamin D, wherein thevitamin D has improved bioavailability when administered to a subject,comprising about 0.1 mg/kg to about 20 mg/kg of MDG based on totalweight of the composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition comprising at least one protein and vitamin D, havingimproved bioavailability of vitamin D, wherein the nutritionalcomposition comprises: an assembly comprising a combination ofmonoglycerides and diglycerides (“MDG”) and vitamin D, wherein thevitamin D has improved bioavailability when administered to a subject,comprising about 0.1 mg/kg to about 20 mg/kg of MDG based on totalweight of the composition, comprising about 0.1 mg/kg to about 10 mg/kgof MDG based on total weight of the composition.

In another aspect, the disclosure may be directed to a nutritionalcomposition comprising at least one protein and vitamin D, havingimproved bioavailability of vitamin D, wherein the nutritionalcomposition comprises: an assembly comprising a combination ofmonoglycerides and diglycerides (“MDG”) and vitamin D, wherein thevitamin D has improved bioavailability when administered to a subject,comprising about 0.1 mg/kg to about 20 mg/kg of MDG based on totalweight of the composition, comprising about 0.1 mg/kg to about 10 mg/kgof MDG based on total weight of the composition, comprising about 10μg/kg to about 30 μg/kg of vitamin D based on total weight of thecomposition.

In another aspect, the disclosure may be directed to a nutritionalcomposition comprising at least one protein and vitamin D, havingimproved bioavailability of vitamin D, wherein the nutritionalcomposition comprises: an assembly comprising a combination ofmonoglycerides and diglycerides (“MDG”) and vitamin D, wherein thevitamin D has improved bioavailability when administered to a subject,comprising about 0.1 mg/kg to about 20 mg/kg of MDG based on totalweight of the composition, comprising about 0.1 mg/kg to about 10 mg/kgof MDG based on total weight of the composition, comprising about 10μg/kg to about 30 μg/kg of vitamin D based on total weight of thecomposition, comprising about 0.1 mg/kg to about 10 mg/kg of MDG andabout 20 μg/kg of vitamin D based on total weight of the composition.

In one aspect, the disclosure may be directed to a nutritionalcomposition having at least one protein, at least one fat, and at leastone lipophilic compound, the composition comprising: an activatedassembly comprising at least one hydrophobic protein, at least onecombination of monoglycerides and diglycerides (“MDG”) and at least onelipophilic compound, wherein at least 15% of the total lipophiliccompound in the nutritional composition remains in the aqueous phaseafter centrifugation at 1,000×g for 1 hour at 20° C.

In one aspect, the disclosure may be directed to a method of preparing anutritional composition having at least one lipophilic compound havingimproved bioavailability, the method comprising the steps of: providinga premix comprising an assembly that comprises at least one hydrophobicprotein, combination of monoglycerides and diglycerides (“MDG”) and atleast one lipophilic compound, wherein the assembly is (i) watersoluble; and (ii) has a size of 100 kD to 1000 kD; adding the premix toan aqueous solution to form an activated premix; and adding theactivated premix to the nutritional composition.

In another aspect, the disclosure may be directed to a method ofpreparing a nutritional composition having at least one lipophiliccompound having improved bioavailability, the method comprising thesteps of: providing a premix comprising an assembly that comprises atleast one hydrophobic protein, combination of monoglycerides anddiglycerides (“MDG”) and at least one lipophilic compound, wherein theassembly is (i) water soluble; and (ii) has a size of 100 kD to 1000 kD;adding the premix to an aqueous solution to form an activated premix;and adding the activated premix to the nutritional composition, whereinthe assembly has a size of 60 kD to 400 kD.

In another aspect, the disclosure may be directed to a method ofpreparing a nutritional composition having at least one lipophiliccompound having improved bioavailability, the method comprising thesteps of: providing a premix comprising an assembly that comprises atleast one hydrophobic protein, combination of monoglycerides anddiglycerides (“MDG”) and at least one lipophilic compound, wherein theassembly is (i) water soluble; and (ii) has a size of 100 kD to 1000 kD;adding the premix to an aqueous solution to form an activated premix;and adding the activated premix to the nutritional composition, whereinthe assembly has a size of 60 kD to 400 kD, wherein the lipophiliccompound is a carotenoid, a lipid soluble vitamin, a lipophilicantioxidant or combinations thereof.

In another aspect, the disclosure may be directed to a method ofpreparing a nutritional composition having at least one lipophiliccompound having improved bioavailability, the method comprising thesteps of: providing a premix comprising an assembly that comprises atleast one hydrophobic protein, combination of monoglycerides anddiglycerides (“MDG”) and at least one lipophilic compound, wherein theassembly is (i) water soluble; and (ii) has a size of 100 kD to 1000 kD;adding the premix to an aqueous solution to form an activated premix;and adding the activated premix to the nutritional composition, whereinthe assembly has a size of 60 kD to 400 kD, wherein the lipophiliccompound is a carotenoid, a lipid soluble vitamin, a lipophilicantioxidant or combinations thereof, wherein the lipophilic compound islutein, vitamin A, vitamin D, vitamin E, vitamin K or any combinationsthereof.

In another aspect, the disclosure may be directed to a method ofpreparing a nutritional composition having at least one lipophiliccompound having improved bioavailability, the method comprising thesteps of: providing a premix comprising an assembly that comprises atleast one hydrophobic protein, combination of monoglycerides anddiglycerides (“MDG”) and at least one lipophilic compound, wherein theassembly is (i) water soluble; and (ii) has a size of 100 kD to 1000 kD;adding the premix to an aqueous solution to form an activated premix;and adding the activated premix to the nutritional composition, whereinthe assembly has a size of 60 kD to 400 kD, wherein the lipophiliccompound is a carotenoid, a lipid soluble vitamin, a lipophilicantioxidant or combinations thereof, wherein the lipophilic compound islutein, vitamin A, vitamin D, vitamin E, vitamin K or any combinationsthereof, wherein the premix is heated to a temperature of 120° F. forabout 10 minutes.

In one aspect, the disclosure may be directed to a method of preparing anutritional composition having at least one lipophilic compound havingimproved bioavailability, the method comprising the steps of: providinga premix comprising an assembly that comprises at least one hydrophobicprotein, a combination of monoglycerides and diglycerides (“MDG”) and atleast one lipophilic compound, wherein the assembly is (i) watersoluble; and (ii) has a size of 100 kD to 1000 kD; adding the premix toa fat-containing solution to form a protected premix; and adding theprotected premix to the nutritional composition.

In another aspect, the disclosure may be directed to a method ofpreparing a nutritional composition having at least one lipophiliccompound having improved bioavailability, the method comprising thesteps of: providing a premix comprising an assembly that comprises atleast one hydrophobic protein, a combination of monoglycerides anddiglycerides (“MDG”) and at least one lipophilic compound, wherein theassembly is (i) water soluble; and (ii) has a size of 100 kD to 1000 kD;adding the premix to a fat-containing solution to form a protectedpremix; and adding the protected premix to the nutritional composition,wherein the assembly has a size of 60 kD to 400 kD.

In another aspect, the disclosure may be directed to a method ofpreparing a nutritional composition having at least one lipophiliccompound having improved bioavailability, the method comprising thesteps of: providing a premix comprising an assembly that comprises atleast one hydrophobic protein, a combination of monoglycerides anddiglycerides (“MDG”) and at least one lipophilic compound, wherein theassembly is (i) water soluble; and (ii) has a size of 100 kD to 1000 kD;adding the premix to a fat-containing solution to form a protectedpremix; and adding the protected premix to the nutritional composition,wherein the assembly has a size of 60 kD to 400 kD, wherein thelipophilic compound is a carotenoid, a lipid soluble vitamin, alipophilic antioxidant or combinations thereof.

In another aspect, the disclosure may be directed to a method ofpreparing a nutritional composition having at least one lipophiliccompound having improved bioavailability, the method comprising thesteps of: providing a premix comprising an assembly that comprises atleast one hydrophobic protein, a combination of monoglycerides anddiglycerides (“MDG”) and at least one lipophilic compound, wherein theassembly is (i) water soluble; and (ii) has a size of 100 kD to 1000 kD;adding the premix to a fat-containing solution to form a protectedpremix; and adding the protected premix to the nutritional composition,wherein the assembly has a size of 60 kD to 400 kD, wherein thelipophilic compound is a carotenoid, a lipid soluble vitamin, alipophilic antioxidant or combinations thereof, wherein the lipophiliccompound is lutein, vitamin A, vitamin D, vitamin E, vitamin K or anycombinations thereof.

In another aspect, the disclosure may be directed to a method ofpreparing a nutritional composition having at least one lipophiliccompound having improved bioavailability, the method comprising thesteps of: providing a premix comprising an assembly that comprises atleast one hydrophobic protein, a combination of monoglycerides anddiglycerides (“MDG”) and at least one lipophilic compound, wherein theassembly is (i) water soluble; and (ii) has a size of 100 kD to 1000 kD;adding the premix to a fat-containing solution to form a protectedpremix; and adding the protected premix to the nutritional composition,wherein the assembly has a size of 60 kD to 400 kD, wherein thelipophilic compound is a carotenoid, a lipid soluble vitamin, alipophilic antioxidant or combinations thereof, wherein the lipophiliccompound is lutein, vitamin A, vitamin D, vitamin E, vitamin K or anycombinations thereof, wherein the premix is heated to a temperature of120° F. for about 10 minutes.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the hourly lymph flow rate over a period of 6 hoursfollowing lutein administration of different nutritional compositionbatches.

FIG. 2 shows the hourly lutein output in lymph over a 6 hour periodafter lutein administration of different nutritional compositionbatches.

FIG. 3 shows cumulative lymphatic lutein absorption over a 6 hour periodafter lutein administration of different nutritional compositionbatches.

FIG. 4 shows the percent change in lymphatic lutein absorption of luteinwith mono- and diglycerides versus control over a period of 6 hours.

FIG. 5 shows the hourly lymph flow rate over a period of 8 hoursfollowing administration of variable nutritional compositions containinglutein.

FIG. 6 shows the hourly lutein output in lymph over an 8 hour periodafter administration of variable nutritional compositions containinglutein.

FIG. 7 shows cumulative lymphatic lutein absorption over a 6 hour and 8hour period after administration of variable nutritional compositionscontaining lutein.

FIG. 8 shows the percent change in lymphatic lutein absorption over aperiod of 6 hours and 8 hours following administration of variablenutritional compositions containing lutein versus control.

FIG. 9 shows the hourly lymphatic triglyceride output over a period of 6hours following nutritional composition administration.

FIG. 10 shows the hourly lymphatic phospholipid output over 6 hoursfollowing nutritional composition administration.

FIG. 11 shows the change in lymphatic absorption of ARA from fasting,over a period of 6 hours following nutritional compositionadministration

FIG. 12 shows the change in lymphatic absorption of DHA from fasting,over a period of 6 hours following nutritional compositionadministration

FIG. 13 shows presence of the high molecular weight lutein assemblywithin the aqueous phase following centrifugations of variablenutritional compositions, as a function of lutein concentration.

FIGS. 14-16 show the effect of different centrifugal forces on thepresence of the FIG. 14: high molecular weight lutein assembly; FIG. 15:protein greater than 137 kD; and FIG. 16: bound lipophile within theaqueous fraction following centrifugation of variable nutritionalcompositions.

FIG. 17 shows the effects that choline and lecithin addition to thenutritional composition have on the formation of the high molecularweight lutein assembly.

FIG. 18 shows the lymphatic output of triglycerides over a period of 6hours following infusion of variable nutritional compositions.

FIG. 19 shows the lymphatic absorption of palmitic acid over a 6 hourperiod following infusion of variable nutritional compositions.

FIG. 20 shows the lymphatic absorption of linoleic acid over a 6 hourperiod following infusion of variable nutritional compositions.

FIG. 21 shows the lymphatic absorption of alpha-linolenic acid over aperiod of 6 hours following infusion of variable nutritionalcompositions.

FIG. 22 shows the change in lymphatic output of DHA over time followingthe administration of variable nutritional compositions.

FIG. 23 shows UV spectra of the aqueous fractions of variablenutritional compositions. UF: ultrafiltration with membrane of 10 kD or100 kD; UC: centrifugation method C (100,000×g); SMP: skimmed milkpowder/nonfat dried milk; WPC: whey protein concentrate 80.

FIG. 24 shows lymphatic triglyceride output over a 6 hour periodfollowing administration of variable nutritional compositions, whereinthe compositions are stratified based on length of time post-production.

DETAILED DESCRIPTION

The present invention relates to a nutritional composition for thedelivery of nutrients, including lipophilic compounds in a more solubleform by using mono- and di-glycerides (hereafter “MDG”) and hydrophobicprotein assembled with the lipophilic protein. This assembly of thenutritional composition containing MDG, hydrophobic protein and alipophilic compound allows for a chaperoning of the lipophilic compoundinto a more soluble form to facilitate easy digestion. The formations ofstable, high molecular weight assemblies (MDG plus hydrophobic proteinand lipophilic protein at approximately 60 kD to approximately 400 kD)enable maximum nutritional delivery and bioavailability of thelipophilic compound due to the increase in these stable, soluble formsover lipophilic compound combinations that do not contain MDG andhydrophobic protein. Increased water soluble forms of the lipophiliccompound lead to increased bioavailability following consumption.

Ultimately, this enables the nutritional composition to deliverlipophilic compounds without over-fortification, which decreasesproduction costs, limits composition complications, and results in amore desirable product. The nutritional composition may be ingested byan infant, toddler, child or adult, thereby providing the nutrientsneeded for proper development, growth, and/or as a nutritionalsupplement. The nutritional composition may further be consumed todeliver lipophilic compounds for the treatment of numerous conditions,including bone health/growth, eye health, cardiovascular health, etc.

2. DEFINITIONS

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art. In case of conflict, the present document, includingdefinitions, will control. Preferred methods and materials are describedbelow, although methods and materials similar or equivalent to thosedescribed herein may be used in practice or testing of the nutritionalcomposition. All publications, patent applications, patents and otherreferences mentioned herein are incorporated by reference in theirentirety. The materials, methods, and examples disclosed herein areillustrative only and not intended to be limiting.

The terms “comprise(s),” “include(s),” “having,” “has,” “can,”“contain(s),” and variants thereof, as used herein, are intended to beopen-ended transitional phrases, terms, or words that do not precludethe possibility of additional acts or structures. The singular forms“a,” “and” and “the” include plural references unless the contextclearly dictates otherwise. The present disclosure also contemplatesother embodiments “comprising,” “consisting of” and “consistingessentially of,” the embodiments or elements presented herein, whetherexplicitly set forth or not.

The term “activated” refers to MDG that has been blended with water orother aqueous liquid. The term “activation” refers to the step ofblending the MDG with the aqueous liquid. For example, an “activated MDGcomponent” means an aqueous liquid into which a MDG has been blended.For the purpose of this disclosure, an activated MDG is typically anaqueous liquid substantially free of oils or fats other than the MDG.

The term “activated assembly” refers to an assembly that comprises atleast one MDG, at least one hydrophobic protein, and at least onelipophilic compound, wherein the assembly may have contacted an aqueousliquid.

The terms “adult formula” and “adult nutritional product” as usedherein, are used interchangeably to refer to formulas for generallymaintaining or improving the health of an adult, and includes thoseformulas designed for adults who have, are susceptible to, or are atrisk of age-related macular degeneration.

The term “bioavailable” as used herein, unless otherwise specified,refers to the ability of a lipophilic compound to be absorbed from thegastrointestinal tract into lymph which will then enter into thebloodstream of an individual such that the substance may be absorbedinto organs and tissues in the body. As the degree of bioavailability ofa compound increases, the compound becomes more likely to enter into andremain in the bloodstream where it may be absorbed and used by the body.As the degree of bioavailability of a compound decreases, the compoundbecomes less likely to being absorbed into lymph from thegastrointestinal tract and would be excreted from the body beforeentering the bloodstream.

The term “casein(s)” as used herein, unless otherwise specified, shouldbe understood to refer to those proteins in bovine milk that willprecipitate from the milk at a solution pH of 4.6 (20° C.); caseinstypically make up about 80% of the protein in whole, undiluted, bovinemilk. Generally, any type of milk-sourced protein source may be utilizedfor the nutritional compositions as a source of bovine beta-casein,including whole milk, nonfat dry milk, milk protein concentrate, totalmilk protein, milk protein isolate, acid casein, calcium caseinate,sodium caseinate, magnesium caseinate, purified beta-casein, andcombinations thereof, all of which will contain beta-casein. Beta-caseincomprises around 30% of the protein contained in cow's milk, and may bepresent as one of several genetic variants, the two most prevalent beinggenetic variants A1 and A2. Bovine beta-casein has further beencategorized into genetic variants including A3, B, C, D, E, F, G, H1,H2, and I.

The term “composition” as used herein, unless otherwise specified,refers to mixtures that are suitable for enteral administration to asubject. Compositions may be in the form of powders, solids,semi-solids, liquids, gels, and semi-liquids. Compositions may furthercomprise vitamins, minerals, and other ingredients.

The term “dry blended” as used herein, unless otherwise specified,refers to the mixing of components or ingredients to form a basenutritional powder or, to the addition of a dry, powdered or granulatedcomponent or ingredient to a base powder to form a powdered nutritionalcomposition.

The terms “fat,” “lipid,” and “oil” as used herein, unless otherwisespecified, are used interchangeably to refer to lipid materials derivedor processed from plants or animals. These terms also include syntheticlipid materials so long as such synthetic materials are suitable foradministration to subjects as defined herein.

The term “glycerides” as used herein refer generally to lipophiliccompounds comprising a glycerol molecule bonded to fatty acid groups.Monoglycerides are glycerol molecules bonded to a single fatty acidgroup; diglycerides are glycerol molecules bonded to two fatty acidgroups; and triglycerides are glycerol molecules bonded to three fattyacid groups. Fats and oils comprise glycerides, and typical fats andoils from animal, fish, algae, vegetable, or seed sources are comprisedprimarily of triglycerides.

The terms “high palmitic acid vegetable oil” or “HPAV oil” refer tovegetable oils comprising greater than about 22% palmitic acid, as apercentage of the total fatty acids in the HPAV oil. Examples of HPAVoil include, but are not limited to, palm oil and palm olein. For thepurpose of this disclosure, HPAV oil may come from sources including,but not limited to, plants, genetically modified plants, fungi, algae,bacteria, and other single-cell organisms. For the purpose of thisdisclosure, the term “HPAV oil” excludes oils and fats that come frommulticellular animal sources, such as beef tallow, chicken fat, fishoil, or milk butterfat.

The term “human milk fortifier” as used herein, unless otherwisespecified, refers to nutritional products suitable for mixing withbreast milk or preterm infant formula or infant formula for consumptionby a preterm or term infant.

The term “infant,” as used herein, unless otherwise specified, refers toa human about 12 months of age or younger. The term “toddler,” as usedherein, unless otherwise specified, refers to a human about 12 months ofage to about 3 years of age. The term “child,” as used herein, unlessotherwise specified, refers to a human about 3 years of age to about 18years of age. The term “adult,” as used herein, unless otherwisespecified, refers to a human about 18 years of age or older.

The terms “infant formula” or “infant nutritional product” as usedherein are used interchangeably to refer to nutritional compositionsthat have the proper balance of macronutrients, micro-nutrients, andcalories to provide sole or supplemental nourishment for and generallymaintain or improve the health of infants, toddlers, or both. Infantformulas preferably comprise nutrients in accordance with the relevantinfant formula guidelines for the targeted consumer or user population,an example of which would be the Infant Formula Act, 21 U.S.C. Section350(a).

The term “lipophilic compound” as used herein refers to components thathave greater solubility in organic solvents such as ethanol, methanol,ethyl ether, acetone, chloroform, benzene, or lipids than they have inwater. Vitamin D is one example of a lipophilic nutrient. For thepurpose of this disclosure, the term “lipophilic nutrient” may beapplied to other lipophilic compounds, including but not limited topharmaceutical compounds.

The terms “monoglyceride and diglyceride oil” or “MDG oil” as usedherein refer to a combination of both monoglycerides and diglyceridesthat are present within a premix. The premix according to the disclosuremay comprise monoglycerides and diglycerides in an amount ofmonoglycerides of at least 12%, including from about 12% to about 98%,and also including from about 20% to about 80%, including from about 20%to about 60%, and including from about 25% to about 50% by weight of thepremix (excluding the weight of the lipophilic compound). The premix maycomprise at least 12%, about 12% to about 98%, about 20% to about 80%,about 20% to about 60% or about 25% to about 50% by weight of themixture of monoglycerides and diglycerides; an amount of lipophiliccompound as described above; and impurities in an amount of less thanabout 20%, less than about 15%, less than about 10%, or less than about2% by weight. It will be recognized by one skilled in the art based onthe disclosure herein that the premix may include a small percentage ofimpurities such that the mixture of monoglycerides and diglycerides isnot 100% by weight solely monoglycerides and diglycerides. In someembodiments, the impurity may be a triglyceride and/or free glycerol.These impurities may typically be less than about 20%, less than about15%, less than about 10%, or less than about 2% by weight. Inembodiments in which the premix comprises monoglycerides and impurities,the balance of the premix (excluding the lipophilic compound) isdiglycerides.

The term “protected premix” refers to a mixture comprising a surfactantand a MDG. The surfactant may be present within, but is not limited to,the premix, the activated premix, and other components of thenutritional composition. Non-limiting examples of suitable surfactantsin a protected premix include lecithin, polyglycerol esters, andcombinations thereof. The protected premix may be made by blending thesurfactant and the MDG in the presence of heat or at ambient temperatureand, in some embodiments, with agitation. The protected premix may beadded to a fat-containing solution to form an “MDG-protected component.”

The term “nutritional composition” as used herein, unless otherwisespecified, refers to nutritional powders, solids, semi-solids, liquids,gels, and semi-liquids that comprise at least one of protein,carbohydrate, and lipid, and are suitable for enteral administration toa subject. The nutritional composition may further comprise vitamins,minerals, and other ingredients, and represent sole, primary, orsupplemental sources of nutrition.

The term “pharmaceutical composition” as used herein, may encompass anysuitable form for dosing to an individual. Non-limiting examples ofdosing means include enteral and parenteral dosage.

The term “powdered nutritional composition” as used herein, unlessotherwise specified, refers to spray dried and/or dry blended powderedand/or agglomerated nutritional compositions comprising a lipid solublenutrient, such as lutein, and a mixture of monoglycerides anddiglycerides, which are reconstitutable with an aqueous liquid, andwhich are suitable for oral administration to a human.

The term “shelf life” as used herein, unless otherwise specified, refersto the stability of the assembly within a, but not limited to, powder,premix, activated premix, protected activated premix, suspension,liquid, concentrated liquid, mixture, or combination thereof.

The term “subject” as used herein refers to a mammal, including but notlimited to a human (e.g., an infant, toddler, child or adult), adomesticated farm animal (e.g., cow, horse, or pig), or a pet (e.g., dogor cat), who ingests the composition.

The terms “susceptible to” or “at risk of” as used herein, unlessotherwise specified, are used interchangeably to mean having littleresistance to a certain condition or disease, including beinggenetically predisposed, having a family history of, and/or havingsymptoms of the condition or disease.

The term “vegetable oil” as used herein refers to a fat derived fromplant, seed, fungal, or algal sources (e.g., not from a multicellularanimal). A vegetable oil may be a solid or liquid fat at roomtemperature. Examples of vegetable oils include, but are not limited to,coconut oil, fractionated coconut oil, soy oil, corn oil, olive oil,safflower oil, high oleic safflower oil, sunflower oil, high oleicsunflower oil, palm oil, palm kernel oil, palm olein, canola oil,cottonseed oil, flax seed oil, hemp seed oil, peanut oil, borage oil,algal oils, fungal oils, and combinations thereof.

To the extent that the terms “includes,” “including,” “contains,” or“containing” are used herein, they are intended to be inclusive in amanner similar to the term “comprising” as that term is interpreted whenemployed as a transitional word in a claim. Furthermore, to the extentthat the term “or” is employed (e.g., A or B) it is intended to mean “Aor B or both.” When the applicants intend to indicate “only A or B butnot both” then the term “only A or B but not both” will be employed.Thus, use of the term “or” herein is the inclusive, and not theexclusive use. Also, to the extent that the terms “in” or “into” areused herein, they are intended to additionally mean “on” or “onto.”

All percentages, parts and ratios as used herein, are by weight of thetotal composition, unless otherwise specified. All such weights as theypertain to listed ingredients are based on the active level and,therefore, do not include solvents or by-products that may be includedin commercially available materials, unless otherwise specified.

All references to singular characteristics or limitations of the presentdisclosure shall include the corresponding plural characteristic orlimitation, and vice versa, unless otherwise specified or clearlyimplied to the contrary by the context in which the reference is made.

All combinations of method or process steps as used herein may beperformed in any order, unless otherwise specified or clearly implied tothe contrary by the context in which the referenced combination is made.

The various embodiments of the nutritional compositions of the presentdisclosure may also be substantially free of any optional or selectedessential ingredient or feature described herein, provided that theremaining composition still contains all of the required ingredients orfeatures as described herein. In this context, and unless otherwisespecified, the term “substantially free” means that the selectedcompositions contain less than a functional amount of the optionalingredient, typically less than 0.1% by weight, and also including zeropercent by weight of such optional or selected essential ingredient.

The nutritional and pharmaceutical compositions and correspondingmanufacturing methods of the present disclosure may comprise, consistof, or consist essentially of the essential elements of the disclosureas described herein, as well as any additional or optional elementdescribed herein or which is otherwise useful in nutritional andpharmaceutical composition applications.

For the recitation of numeric ranges herein, each intervening numberthere between with the same degree of precision is explicitlycontemplated. For example, for the range of 6-9, the numbers 7 and 8 arecontemplated in addition to 6 and 9, and for the range 6.0-7.0, thenumber 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 areexplicitly contemplated.

3. NUTRIENT COMPOSITION

Provided herein is a nutritional composition. The nutritionalcomposition comprises at least one protein, at least one fat, and atleast one lipophilic compound. As explained below, the nutritionalcomposition includes assemblies of agglomerated compounds that increasethe bioavailability of the lipophilic compounds following consumption.The nutritional composition may include an assembly comprising at leastone lipophilic compound, at least one protein, and at least one fat. Thecombination of fat (e.g., MDG) and protein (e.g., hydrophobic protein)interacts with lipophilic compounds, such as lutein and Vitamin D,enhancing solubility and stability of lipophilic compounds, compared toassemblies without MDG and hydrophobic protein. The nutritionalcomposition may comprise a premix, an activated premix, and a protectedpremix.

The nutritional composition may be used for consumption. The nutritionalcomposition may be ingested by an infant and thus, provides the infantnutrients needed for proper development and growth. The nutritionalcomposition may also be ingested by a toddler or child, for properdelivery of nutrients for continued development and growth. Thenutritional composition may also be ingested by an adult, as anutritional supplement. The nutritional composition can be used todeliver lipophilic compounds for the treatment of numerous conditions,including bone health/growth, eye health, cardiovascular health, etc.

The composition may be in any useful form. Non-limiting product formsinclude: solids, semi-solids, ready-to-drink liquids, concentratedliquids, gels, and powders. In some embodiments, the composition may bein the form of a flowable or substantially flowable particulatecomposition. In some embodiments, the composition may be easily scoopedand measured with a spoon or similar other device, such that thecomposition may easily be reconstituted by the intended user with asuitable aqueous liquid, such as water, to form a liquid nutritional orpharmaceutical composition for immediate oral or enteral use. In thiscontext, “immediate” use generally means within about 48 hours, mosttypically within about 24 hours, and in some embodiments, immediatelyafter reconstitution.

In some embodiments, the composition may include spray dried powders,dry blended powders, agglomerated powders, combinations thereof, orpowders prepared by other suitable methods.

In some embodiments, the composition may be contained in a dosageelement suitable for oral consumption. Suitable dosage elements includetablets, hard gelatin capsules, starch capsules, cellulose-basedcapsules, softgel capsules, and elixirs.

In some embodiments, the powdered composition may be compressed into atablet. In some embodiments, the powdered composition may be included ina capsule. Capsules comprise a shell surrounding and containing thecomposition. The capsule shell dissolves or disintegrates in thedigestive tract after the capsule is ingested, thereby releasing thecomposition to be absorbed by the body of the individual ingesting thecapsule. Capsule shells are typically made of gelatin, starch,cellulose, or other components that readily dissolve or disintegrateafter ingestion, and the composition, manufacture, and use of capsuleshells are well known in the art. In some embodiments, the dosageelement for the composition is a softgel capsule. Softgel capsules areparticularly suitable for containing liquid-based ingredients, such asnutrients dissolved, dispersed or suspended in a carrier oil. The shellof a softgel capsule is typically made of gelatin plasticized withglycerin and water, although vegetarian softgel capsules made fromstarch or carrageenan are also available. Softgel capsule shells aretypically made and filled with the composition in continuous processesthat are known in the art. Softgel capsules are made by manufacturerssuch as Catalent Pharma Solutions, LLC (Somerset, N.J.) and CaptekSoftgel International (Cerritos, Calif.).

Capsules come in a wide range of sizes, and the capsule size should bechosen to contain an appropriate volume or weight of the compositionand, hence, an appropriate dosage of the lipophilic compound. Thecapsule size can be chosen to contain at least about 0.1 grams of thecomposition, including from about 0.1 grams to about 30 grams, fromabout 0.2 grams to about 20 grams, from about 0.25 grams to about 15grams, from about 0.25 grams to about 10 grams, from about 0.25 grams toabout 5 grams, from about 0.25 to about 3 grams, from about 0.25 gramsto about 1.5 grams, or about 0.25 to about 1.0 grams of the composition.In some embodiments, for use with humans, the capsules contain fromabout 0.1 grams to about 1.5 grams or from about 0.2 grams to about 1.0grams of the composition, as these capsule sizes are most convenient formost adults and children to swallow. In some embodiments, for use withlarge animals, such as domesticated farm animals, the capsules containfrom about 0.1 grams to about 30 grams or from about 1.0 grams to about30 grams of the composition.

The nutritional composition may be formulated with sufficient kinds andamounts of nutrients so as to provide a sole, primary, or supplementalsource of nutrition, or to provide a specialized nutritional compositionfor use in individuals afflicted with specific diseases or conditions.

The nutritional composition may have a caloric density tailored to thenutritional needs of the ultimate user, although in most instances thenutritional composition may comprise from about 65 to about 800 kcal/240mL. The nutritional composition, as discussed herein, provides a methodto easily and effectively control caloric intake to an individual (e.g.,infant, toddler, child or adult). The ability to tightly control caloricintake is important because different individuals have different caloricneeds. The nutritional composition may comprise a caloric density ofabout 75 to about 700 kcal/240 mL. The nutritional composition maycomprise a caloric density of about 100 to about 650 kcal/240 mL. Forexample, the nutritional composition may comprise about 65 kcal/240 mL,70 kcal/240 mL, 75 kcal/240 mL, 80 kcal/240 mL, 85 kcal/240 mL, 90kcal/240 mL, 95 kcal/240 mL, 100 kcal/240 mL, 105 kcal/240 mL, 110kcal/240 mL, 115 kcal/240 mL, 120 kcal/240 mL, 125 kcal/240 mL, 130kcal/240 mL, 135 kcal/240 mL, 140 kcal/240 mL, 145 kcal/240 mL, 150kcal/240 mL, 155 kcal/240 mL, 160 kcal/240 mL, 165 kcal/240 mL, 170kcal/240 mL, 175 kcal/240 mL, 180 kcal/240 mL, 185 kcal/240 mL, 190kcal/240 mL, 195 kcal/240 mL, 200 kcal/240 mL, 205 kcal/240 mL, 210kcal/240 mL, 215 kcal/240 mL, 220 kcal/240 mL, 225 kcal/240 mL, 230kcal/240 mL, 235 kcal/240 mL, 240 kcal/240 mL, 245 kcal/240 mL, 250kcal/240 mL, 255 kcal/240 mL, 260 kcal/240 mL, 265 kcal/240 mL, 270kcal/240 mL, 275 kcal/240 mL, 280 kcal/240 mL, 285 kcal/240 mL, 290kcal/240 mL, 295 kcal/240 mL, 300 kcal/240 mL, 305 kcal/240 mL, 310kcal/240 mL, 315 kcal/240 mL, 320 kcal/240 mL, 325 kcal/240 mL, 330kcal/240 mL, 335 kcal/240 mL, 340 kcal/240 mL, 345 kcal/240 mL, 350kcal/240 mL, 355 kcal/240 mL, 360 kcal/240 mL, 365 kcal/240 mL, 370kcal/240 mL, 375 kcal/240 mL, 380 kcal/240 mL, 385 kcal/240 mL, 390kcal/240 mL, 395 kcal/240 mL, 400 kcal/240 mL, 405 kcal/240 mL, 410kcal/240 mL, 415 kcal/240 mL, 420 kcal/240 mL, 425 kcal/240 mL, 430kcal/240 mL, 435 kcal/240 mL, 440 kcal/240 mL, 445 kcal/240 mL, 450kcal/240 mL, 455 kcal/240 mL, 460 kcal/240 mL, 465 kcal/240 mL, 470kcal/240 mL, 475 kcal/240 mL, 480 kcal/240 mL, 485 kcal/240 mL, 490kcal/240 mL, 495 kcal/240 mL, 500 kcal/240 mL, 550 kcal/240 mL, 600kcal/240 mL, 650 kcal/240 mL, 700 kcal/240 mL, 750 kcal/240 mL, or 800kcal/240 mL.

Further, non-limiting examples of the composition include human milkfortifiers, preterm infant formulas, infant formulas, elemental andsemi-elemental formulas, pediatric formulas, toddler formulas, adultformulas, nutritional supplements, capsules, suppositories, sprays,drops, lotions, ointments, microcapsules, and liposomes.

The at least one lipophilic compound may be in the nutritionalcomposition at about 1 μg/kg to about 10 g/kg of lipophilic compound.The at least one lipophilic compound may be in the nutritionalcomposition at about 1 μg/kg to about 7 g/kg of lipophilic compound. Theat least one lipophilic compound may be in the nutritional compositionat about 500 μg/kg to about 5 g/kg of lipophilic compound. The at leastone lipophilic compound may be in the nutritional composition at about 1mg/kg to about 1 g/kg of lipophilic compound. The at least onelipophilic compound may be in the nutritional composition at about 100mg/kg to about 1 g/kg of lipophilic compound. For example, thelipophilic compound may be in the nutritional composition at about 1μg/kg, 10 μg/kg, 50 μg/kg, 100 μg/kg, 150 μg/kg, 200 μg/kg, 250 μg/kg,300 μg/kg, 350 μg/kg, 400 μg/kg, 450 μg/kg, 500 μg/kg, 550 μg/kg, 600μg/kg, 650 μg/kg, 700 μg/kg, 750 μg/kg, 800 μg/kg, 850 μg/kg, 900 μg/kg,950 μg/kg, 1 mg/kg, 10 mg/kg, 50 mg/kg, 100 mg/kg, 150 mg/kg, 200 mg/kg,250 mg/kg, 300 mg/kg, 350 mg/kg, 400 mg/kg, 450 mg/kg, 500 mg/kg, 550mg/kg, 600 mg/kg, 650 mg/kg, 700 mg/kg, 750 mg/kg, 800 mg/kg, 850 mg/kg,900 mg/kg, 950 mg/kg, 1 g/kg, 0.5 g/kg, 1 g/kg, 1.5 g/kg, 2 g/kg, 2.5g/kg, 3 g/kg, 3.5 g/kg, 4 g/kg, 4.5 g/kg, 5 g/kg, 5.5 g/kg, 6 g/kg, 6.5g/kg, 7 g/kg, 7.5 g/kg, 8 g/kg, 8.5 g/kg, 9 g/kg, 9.5 g/kg, or 10 g/kg.

The at least one fat may be in the nutritional composition at about 10g/kg to about 100 g/kg. The at least one fat may be in the nutritionalcomposition at about 20 g/kg to about 80 g/kg. The at least one fat maybe in the nutritional composition at about 30 g/kg to about 70 g/kg. Theat least one fat may be in the nutritional composition at about 40 g/kgto about 60 g/kg. For example, the at least one fat may be in thenutritional composition at about 10 g/kg, 10.5 g/kg, 11 g/kg, 11.5 g/kg,12 g/kg, 12.5 g/kg, 13 g/kg, 13.5 g/kg, 14 g/kg, 14.5 g/kg, 15 g/kg,15.5 g/kg, 16 g/kg, 16.5 g/kg, 17 g/kg, 17.5 g/kg, 18 g/kg, 18.5 g/kg,19 g/kg, 19.5 g/kg, 20 g/kg, 20.5 g/kg, 21 g/kg, 21.5 g/kg, 22 g/kg,22.5 g/kg, 23 g/kg, 23.5 g/kg, 24 g/kg, 24.5 g/kg, 25 g/kg, 25.5 g/kg,26 g/kg, 26.5 g/kg, 27 g/kg, 27.5 g/kg, 28 g/kg, 28.5 g/kg, 29 g/kg,29.5 g/kg, 30 g/kg, 30.5 g/kg, 31 g/kg, 31.5 g/kg, 32 g/kg, 32.5 g/kg,33 g/kg, 33.5 g/kg, 34 g/kg, 34.5 g/kg, 35 g/kg, 35.5 g/kg, 36 g/kg,36.5 g/kg, 37 g/kg, 37.5 g/kg, 38 g/kg, 38.5 g/kg, 39 g/kg, 39.5 g/kg,40 g/kg, 40.5 g/kg, 41 g/kg, 41.5 g/kg, 42 g/kg, 42.5 g/kg, 43 g/kg,43.5 g/kg, 44 g/kg, 44.5 g/kg, 45 g/kg, 45.5 g/kg, 46 g/kg, 46.5 g/kg,47 g/kg, 47.5 g/kg, 48 g/kg, 48.5 g/kg, 49 g/kg, 49.5 g/kg, 50 g/kg,50.5 g/kg, 51 g/kg, 51.5 g/kg, 52 g/kg, 52.5 g/kg, 53 g/kg, 53.5 g/kg,54 g/kg, 54.5 g/kg, 55 g/kg, 55.5 g/kg, 56 g/kg, 56.5 g/kg, 57 g/kg,57.5 g/kg, 58 g/kg, 58.5 g/kg, 59 g/kg, 59.5 g/kg, 60 g/kg, 60.5 g/kg,61 g/kg, 61.5 g/kg, 62 g/kg, 62.5 g/kg, 63 g/kg, 63.5 g/kg, 64 g/kg,64.5 g/kg, 65 g/kg, 65.5 g/kg, 66 g/kg, 66.5 g/kg, 67 g/kg, 67.5 g/kg,68 g/kg, 68.5 g/kg, 69 g/kg, 69.5 g/kg, 70 g/kg, 70.5 g/kg, 71 g/kg,71.5 g/kg, 72 g/kg, 72.5 g/kg, 73 g/kg, 73.5 g/kg, 74 g/kg, 74.5 g/kg,75 g/kg, 75.5 g/kg, 76 g/kg, 76.5 g/kg, 77 g/kg, 77.5 g/kg, 78 g/kg,78.5 g/kg, 79 g/kg, 79.5 g/kg, 80 g/kg, 80.5 g/kg, 81 g/kg, 81.5 g/kg,82 g/kg, 82.5 g/kg, 83 g/kg, 83.5 g/kg, 84 g/kg, 84.5 g/kg, 85 g/kg,85.5 g/kg, 86 g/kg, 86.5 g/kg, 87 g/kg, 87.5 g/kg, 88 g/kg, 88.5 g/kg,89 g/kg, 89.5 g/kg, 90 g/kg, 90.5 g/kg, 91 g/kg, 91.5 g/kg, 92 g/kg,92.5 g/kg, 93 g/kg, 93.5 g/kg, 94 g/kg, 94.5 g/kg, 95 g/kg, 95.5 g/kg,96 g/kg, 96.5 g/kg, 97 g/kg, 97.5 g/kg, 98 g/kg, 98.5 g/kg, 99 g/kg,99.5 g/kg, or 100 g/kg.

The at least one protein may be in the nutritional composition at about10 g/kg to about 100 g/kg. The at least one protein may be in thenutritional composition at about 20 g/kg to about 80 g/kg. The at leastone protein may be in the nutritional composition at about 30 g/kg toabout 70 g/kg. The at least one protein may be in the nutritionalcomposition at about 40 g/kg to about 60 g/kg. For example, the at leastone protein may be in the nutritional composition at about 10 g/kg, 10.5g/kg, 11 g/kg, 11.5 g/kg, 12 g/kg, 12.5 g/kg, 13 g/kg, 13.5 g/kg, 14g/kg, 14.5 g/kg, 15 g/kg, 15.5 g/kg, 16 g/kg, 16.5 g/kg, 17 g/kg, 17.5g/kg, 18 g/kg, 18.5 g/kg, 19 g/kg, 19.5 g/kg, 20 g/kg, 20.5 g/kg, 21g/kg, 21.5 g/kg, 22 g/kg, 22.5 g/kg, 23 g/kg, 23.5 g/kg, 24 g/kg, 24.5g/kg, 25 g/kg, 25.5 g/kg, 26 g/kg, 26.5 g/kg, 27 g/kg, 27.5 g/kg, 28g/kg, 28.5 g/kg, 29 g/kg, 29.5 g/kg, 30 g/kg, 30.5 g/kg, 31 g/kg, 31.5g/kg, 32 g/kg, 32.5 g/kg, 33 g/kg, 33.5 g/kg, 34 g/kg, 34.5 g/kg, 35g/kg, 35.5 g/kg, 36 g/kg, 36.5 g/kg, 37 g/kg, 37.5 g/kg, 38 g/kg, 38.5g/kg, 39 g/kg, 39.5 g/kg, 40 g/kg, 40.5 g/kg, 41 g/kg, 41.5 g/kg, 42g/kg, 42.5 g/kg, 43 g/kg, 43.5 g/kg, 44 g/kg, 44.5 g/kg, 45 g/kg, 45.5g/kg, 46 g/kg, 46.5 g/kg, 47 g/kg, 47.5 g/kg, 48 g/kg, 48.5 g/kg, 49g/kg, 49.5 g/kg, 50 g/kg, 50.5 g/kg, 51 g/kg, 51.5 g/kg, 52 g/kg, 52.5g/kg, 53 g/kg, 53.5 g/kg, 54 g/kg, 54.5 g/kg, 55 g/kg, 55.5 g/kg, 56g/kg, 56.5 g/kg, 57 g/kg, 57.5 g/kg, 58 g/kg, 58.5 g/kg, 59 g/kg, 59.5g/kg, 60 g/kg, 60.5 g/kg, 61 g/kg, 61.5 g/kg, 62 g/kg, 62.5 g/kg, 63g/kg, 63.5 g/kg, 64 g/kg, 64.5 g/kg, 65 g/kg, 65.5 g/kg, 66 g/kg, 66.5g/kg, 67 g/kg, 67.5 g/kg, 68 g/kg, 68.5 g/kg, 69 g/kg, 69.5 g/kg, 70g/kg, 70.5 g/kg, 71 g/kg, 71.5 g/kg, 72 g/kg, 72.5 g/kg, 73 g/kg, 73.5g/kg, 74 g/kg, 74.5 g/kg, 75 g/kg, 75.5 g/kg, 76 g/kg, 76.5 g/kg, 77g/kg, 77.5 g/kg, 78 g/kg, 78.5 g/kg, 79 g/kg, 79.5 g/kg, 80 g/kg, 80.5g/kg, 81 g/kg, 81.5 g/kg, 82 g/kg, 82.5 g/kg, 83 g/kg, 83.5 g/kg, 84g/kg, 84.5 g/kg, 85 g/kg, 85.5 g/kg, 86 g/kg, 86.5 g/kg, 87 g/kg, 87.5g/kg, 88 g/kg, 88.5 g/kg, 89 g/kg, 89.5 g/kg, 90 g/kg, 90.5 g/kg, 91g/kg, 91.5 g/kg, 92 g/kg, 92.5 g/kg, 93 g/kg, 93.5 g/kg, 94 g/kg, 94.5g/kg, 95 g/kg, 95.5 g/kg, 96 g/kg, 96.5 g/kg, 97 g/kg, 97.5 g/kg, 98g/kg, 98.5 g/kg, 99 g/kg, 99.5 g/kg, or 100 g/kg.

The ratios and amounts between the different compounds are critical inthe resultant assemblies formed during composition. In order to formstable, high molecular weight assemblies, the w/w ratio of MDG tolipophilic compound may be in the nutritional composition at about12,000:1 to about 1:1. The w/w ratio of MDG to lipophilic compound maybe in the nutritional composition at about 10,000:1 to about 1:1. Thew/w ratio of MDG to lipophilic compound may be in the nutritionalcomposition at about 1,000:1 to about 1:1. The w/w ratio of MDG tolipophilic compound may be in the nutritional composition at about 100:1to about 1:1. For example, w/w ratio of MDG to lipophilic compound maybe in the nutritional composition at about 12000:1, 11500:1, 11000:1,10500:1, 10000:1, 9500:1, 9000:1, 8500:1, 8000:1, 7500:1, 7000:1,6500:1, 6000:1, 5500:1, 5000:1, 4900:1, 4800:1, 4700:1, 4600:1, 4500:1,4400:1, 4300:1, 4200:1, 4100:1, 4000:1, 3900:1, 3800:1, 3700:1, 3600:1,3500:1, 3400:1, 3300:1, 3200:1, 3100:1, 3000:1, 2900:1, 2800:1, 2700:1,2600:1, 2500:1, 2400:1, 2300:1, 2200:1, 2100:1, 2000:1, 1900:1, 1800:1,1700:1, 1600:1, 1500:1, 1400:1, 1300:1, 1200:1, 1100:1, 1000:1, 950:1,900:1, 850:1, 800:1, 750:1, 700:1, 650:1, 600:1, 550:1, 500:1, 450:1,400:1, 350:1, 300:1, 250:1, 200:1, 150:1, 100:1, 90:1, 80:1, 70:1, 60:1,50:1, 40:1, 30:1, 20:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, or1:1.

One embodiment of the nutritional composition may comprise about 145mg/kg to about 600 mg/kg of MDG and about 1.12 g/kg of a lipophiliccompound.

In another embodiment, the nutritional composition may comprise about0.1 mg/kg to about 10 mg/kg of MDG and about 20 μg/kg of vitamin D.

b. Assembly

The nutritional composition may comprise an assembly. The assembly maycomprise at least one fat, at least one protein, and at least onelipophilic compound. The assembly may comprise MDG, at least onehydrophobic protein, and at least one lipophilic compound. It is wellknown in the art that lipophilic compounds have limited aqueoussolubility, resulting in their decreased bioavailability when consumedas part of a nutritional composition. The assembly increases the aqueoussolubility of lipophilic compounds. Accordingly, the assembly increasesthe bioavailability of the lipophilic compounds, and their resultantabsorption within the gastrointestinal (GI) tract, relative to controlswithout MDG and hydrophobic protein.

The assembly is water soluble, and the interactions between MDG, the atleast one hydrophobic protein and at least one lipophilic compound allowfor stability of the assembly to withstand high-speed centrifugalforces. For example, the nutritional composition may be centrifuged at1×g, 50×g, 100×g, 500×g, 1000×g, 1500×g, 2000×g, 2500×g, 3000×g, 3500×g,4000×g, 4500×g, 5000×g, 5500×g, 6000×g, 6500×g, 7000×g, 7500×g, 8000×g,8500×g, 9000×g, 9500×g, 10000×g, 10500×g, 11000×g, 11500×g, 12000×g,12500×g, 13000×g, 13500×g, 14000×g, 14500×g, 15000×g, 15500×g, 16000×g,16500×g, 17000×g, 17500×g, 18000×g, 18500×g, 19000×g, 19500×g, 20000×g,20500×g, 21000×g, 21500×g, 22000×g, 22500×g, 23000×g, 23500×g, 24000×g,24500×g, 25000×g, 25500×g, 26000×g, 26500×g, 27000×g, 27500×g, 28000×g,28500×g, 29000×g, 29500×g, 30000×g, 30500×g, 31000×g, 31500×g, 32000×g,32500×g, 33000×g, 33500×g, 34000×g, 34500×g, 35000×g, 35500×g, 36000×g,36500×g, 37000×g, 37500×g, 38000×g, 38500×g, 39000×g, 39500×g, 40000×g,40500×g, 41000×g, 41500×g, 42000×g, 42500×g, 43000×g, 43500×g, 44000×g,44500×g, 45000×g, 45500×g, 46000×g, 46500×g, 47000×g, 47500×g, 48000×g,48500×g, 49000×g, 49500×g, 50000×g, 50500×g, 51000×g, 51500×g, 52000×g,52500×g, 53000×g, 53500×g, 54000×g, 54500×g, 55000×g, 55500×g, 56000×g,56500×g, 57000×g, 57500×g, 58000×g, 58500×g, 59000×g, 59500×g, 60000×g,60500×g, 61000×g, 61500×g, 62000×g, 62500×g, 63000×g, 63500×g, 64000×g,64500×g, 65000×g, 65500×g, 66000×g, 66500×g, 67000×g, 67500×g, 68000×g,68500×g, 69000×g, 69500×g, 70000×g, 70500×g, 71000×g, 71500×g, 72000×g,72500×g, 73000×g, 73500×g, 74000×g, 74500×g, 75000×g, 75500×g, 76000×g,76500×g, 77000×g, 77500×g, 78000×g, 78500×g, 79000×g, 79500×g, 80000×g,80500×g, 81000×g, 81500×g, 82000×g, 82500×g, 83000×g, 83500×g, 84000×g,84500×g, 85000×g, 85500×g, 86000×g, 86500×g, 87000×g, 87500×g, 88000×g,88500×g, 89000×g, 89500×g, 90000×g, 90500×g, 91000×g, 91500×g, 92000×g,92500×g, 93000×g, 93500×g, 94000×g, 94500×g, 95000×g, 95500×g, 96000×g,96500×g, 97000×g, 97500×g, 98000×g, 98500×g, 99000×g, 99500×g, or100000×g.

In some embodiments, the assembly is still water soluble aftercentrifugation at about 31,000×g for at least 1 hour at 20° C. In someembodiments, at least 5% of the assembly remains in the aqueous phaseafter centrifugation.

The nutritional composition may be spun down using a centrifuge toassess the amount of fat, protein and lipophilic compound remaining inthe aqueous phase. It is hypothesized, without being bound to anyparticular theory, that the fat and lipophilic compound that remain inthe aqueous phase following centrifugation are likely associated withthe assembly due to their limited aqueous solubility. It should be notedthat different centrifugal speeds and times may be used to spin down thenutritional composition. Accordingly, one skilled in the art wouldappreciate that different centrifugal conditions may provide differentamounts of the assembly, and corresponding compounds, within the aqueousphase, but will still allow quantification of desired compound.

In some embodiments, the details of centrifugation are as follows: speedof centrifugation: 1000×g, centrifugation time: 15 min, centrifugationtemperature: 20° C., centrifuge tubes: Cellstar tube 50 ml from Greinerbio-one, tube diameter: 23 mm, amount of product in tube: 45 ml,centrifuge: type 5810 R from Eppendorf, and aqueous phase isolation bysyringe through the cream layer.

In some embodiments, the details of centrifugation are as follows: speedof centrifugation: 4500×g, centrifugation time: 15 min, centrifugationtemperature: 20° C., centrifuge tubes: Cellstar tube 50 ml from Greinerbio-one, tube diameter: 23 mm, amount of product in tube: 45 ml,centrifuge: type 5810 R from Eppendorf, and aqueous phase isolation bysyringe through the cream layer.

In some embodiments, the details of centrifugation are as follows: speedof centrifugation: 100,000×g, centrifugation time: 1 h, centrifugationtemperature: 20° C., centrifuge tubes: Ultra Clear Centrifuge tubes fromBeckmann Coulter, tube size: 14×89 mm, amount of product in tube: 11.3g, centrifuge: type L-90K from Beckmann Coulter, aqueous phase isolationby freezing the tube with content in liquid nitrogen and cutting thepart with the serum phase including the tube wall followed by thawingthe content of this part of the tube.

In some embodiments, the centrifuge procedure may be performed asfollows, approximately 35 g of nutritional composition is transferredinto a 50-mL polypropylene centrifuge tube (capable of withstanding100,000×g; e.g., VWR catalog no. 21007-290). The tubes are capped andplaced in a JA-20 rotor in a Beckman Coulter Model Avanti J-Ecentrifuge. The temperature is set at 20° C., and the tubes arecentrifuged at 31,000×g (16,000 rpm) for 1 to 4 hours. The tubes areremoved from the centrifuge, and approximately 0.6 to 1.0 g ofsupernatant is transferred into a tared 10 mL volumetric flask, and thesupernatant sample weight is recorded. The supernatant sample in the 10mL volumetric flask is diluted to 7.0 g with Milli-Q Plus water, andstirred for five minutes. The diluted supernatant sample in the 10 mLvolumetric flask is then diluted to 10 mL with acetonitrile containingtrifluoroacetic acid at 0.33% (v/v), and stirred for an additional fiveminutes. The flask is stoppered, and inverted repeatedly to thoroughlymix. A 1.0 to 1.5 mL aliquot is transferred from the flask into an HPLCautosampler vial. The vial is capped, and the aliquot is tested for thepresence of different variables within the aqueous phase, such as MDG,lutein, hydrophobic protein, etc.

In one embodiment, the nutritional composition may be centrifuged at1,000×g for 1 hour at 20° C. In one embodiment, the nutritionalcomposition may be centrifuged at 31,000×g for 1 hour at 20° C. In oneembodiment, the nutritional composition may be centrifuged at 100,000×gfor 1 hour at 20° C.

MDG may be in the aqueous phase at about 0.001% to about 50% of thenutritional composition, following centrifugation at 1,000×g for 1 hourat 20° C. MDG may be in the aqueous phase at about 0.5% to about 40% ofthe nutritional composition, following centrifugation at 1,000×g for 1hour at 20° C. MDG may be in the aqueous phase at about 5% to about 30%of the nutritional composition, following centrifugation at 1,000×g for1 hour at 20° C. MDG may be in the aqueous phase at about 15% to about25% of the nutritional composition, following centrifugation at 1,000×gfor 1 hour at 20° C. For example, MDG may be in the aqueous phase atabout 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%,0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%,1.25%, 1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%,4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%,7.25%, 7.5%, 7.75%, 8%, 8.25%, 8.5%, 8.75%, 9%, 9.25%, 9.5%, 9.75%, 10%,10.25%, 10.5%, 10.75%, 11%, 11.25%, 11.5%, 11.75%, 12%, 12.25%, 12.5%,12.75%, 13%, 13.25%, 13.5%, 13.75%, 14%, 14.25%, 14.5%, 14.75%, 15%,15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%, 17.5%,17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%, 19.75%, 20%,20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%,22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%,25.25%, 25.5%, 25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%,27.75%, 28%, 28.25%, 28.5%, 28.75%, 29%, 29.25%, 29.5%, 29.75%, 30%,30.25%, 30.5%, 30.75%, 31%, 31.25%, 31.5%, 31.75%, 32%, 32.25%, 32.5%,32.75%, 33%, 33.25%, 33.5%, 33.75%, 34%, 34.25%, 34.5%, 34.75%, 35%,35.25%, 35.5%, 35.75%, 36%, 36.25%, 36.5%, 36.75%, 37%, 37.25%, 37.5%,37.75%, 38%, 38.25%, 38.5%, 38.75%, 39%, 39.25%, 39.5%, 39.75%, 40%,40.25%, 40.5%, 40.75%, 41%, 41.25%, 41.5%, 41.75%, 42%, 42.25%, 42.5%,42.75%, 43%, 43.25%, 43.5%, 43.75%, 44%, 44.25%, 44.5%, 44.75%, 45%,45.25%, 45.5%, 45.75%, 46%, 46.25%, 46.5%, 46.75%, 47%, 47.25%, 47.5%,47.75%, 48%, 48.25%, 48.5%, 48.75%, 49%, 49.25%, 49.5%, 49.75%, or 50%of the nutritional composition, following centrifugation at 1,000×g for1 hour at 20° C.

MDG may be in the aqueous phase at about 0.001% to about 30% of thenutritional composition, following centrifugation at 31,000×g for 1 hourat 20° C. MDG may be in the aqueous phase at about 0.1% to about 20% ofthe nutritional composition, following centrifugation at 31,000×g for 1hour at 20° C. MDG may be in the aqueous phase at about 1% to about 5%of the nutritional composition, following centrifugation at 31,000×g for1 hour at 20° C. For example, MDG may be in the aqueous phase at about0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%,0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%,1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%,4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%,7.5%, 7.75%, 8%, 8.25%, 8.5%, 8.75%, 9%, 9.25%, 9.5%, 9.75%, 10%,10.25%, 10.5%, 10.75%, 11%, 11.25%, 11.5%, 11.75%, 12%, 12.25%, 12.5%,12.75%, 13%, 13.25%, 13.5%, 13.75%, 14%, 14.25%, 14.5%, 14.75%, 15%,15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%, 17.5%,17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%, 19.75%, 20%,20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%,22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%,25.25%, 25.5%, 25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%,27.75%, 28%, 28.25%, 28.5%, 28.75%, 29%, 29.25%, 29.5%, 29.75%, or 30%of the nutritional composition, following centrifugation at 31,000×g for1 hour at 20° C.

MDG may be in the aqueous phase at about 0.001% to about 20% of thenutritional composition, following centrifugation at 100,000×g for 1hour at 20° C. MDG may be in the aqueous phase at about 0.01% to about15% of the nutritional composition, following centrifugation at100,000×g for 1 hour at 20° C. MDG may be in the aqueous phase at about0.1% to about 10% of the nutritional composition, followingcentrifugation at 100,000×g for 1 hour at 20° C. MDG may be in theaqueous phase at about 1% to about 5% of the nutritional composition,following centrifugation at 100,000×g for 1 hour at 20° C. For example,MDG may be in the aqueous phase at about 0.001%, 0.05%, 0.1%, 0.15%,0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%,0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%,1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%,2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%,4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%,5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%,6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%,7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%,8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%,10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%,11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12%,12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%, 12.7%, 12.8%, 12.9%, 13%,13.1%, 13.2%, 13.3%, 13.4%, 13.5%, 13.6%, 13.7%, 13.8%, 13.9%, 14%,14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%, 14.7%, 14.8%, 14.9%, 15%,15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%,16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%,17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%,18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%,19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20% ofthe nutritional composition, following centrifugation at 100,000×g for 1hour at 20° C.

MDG may be in the assembly at about 0.001% to about 20% of thenutritional composition, following centrifugation at 31,000×g for 1 hourat 20° C. MDG may be in the assembly at about 0.01% to about 15% of thenutritional composition, following centrifugation at 31,000×g for 1 hourat 20° C. MDG may be in the assembly at about 0.1% to about 10% of thenutritional composition, following centrifugation at 31,000×g for 1 hourat 20° C. MDG may be in the assembly at about 1% to about 5% of thenutritional composition, following centrifugation at 31,000×g for 1 hourat 20° C. For example, MDG may be in the assembly at about 0.001%,0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%,0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%,1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%,2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%,3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%,4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%,6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%,7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%,8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%,9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%,10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%,11.8%, 11.9%, 12%, 12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%, 12.7%,12.8%, 12.9%, 13%, 13.1%, 13.2%, 13.3%, 13.4%, 13.5%, 13.6%, 13.7%,13.8%, 13.9%, 14%, 14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%, 14.7%,14.8%, 14.9%, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%,15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%,16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%,17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%,18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%,19.8%, 19.9%, or 20% of the nutritional composition, followingcentrifugation at 31,000×g for 1 hour at 20° C.

Lutein, a lipophilic compound discussed below, may be in the aqueousphase at about 0.001% to about 50% of the nutritional composition,following centrifugation at 1,000×g for 1 hour at 20° C. Lutein may bein the aqueous phase at about 0.5% to about 40% of the nutritionalcomposition, following centrifugation at 1,000×g for 1 hour at 20° C.Lutein may be in the aqueous phase at about 5% to about 30% of thenutritional composition, following centrifugation at 1,000×g for 1 hourat 20° C. Lutein may be in the aqueous phase at about 15% to about 25%of the nutritional composition, following centrifugation at 1,000×g for1 hour at 20° C. For example, lutein may be in the aqueous phase atabout 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%,0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%,1.25%, 1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%,4.25%, 4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%,7.25%, 7.5%, 7.75%, 8%, 8.25%, 8.5%, 8.75%, 9%, 9.25%, 9.5%, 9.75%, 10%,10.25%, 10.5%, 10.75%, 11%, 11.25%, 11.5%, 11.75%, 12%, 12.25%, 12.5%,12.75%, 13%, 13.25%, 13.5%, 13.75%, 14%, 14.25%, 14.5%, 14.75%, 15%,15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%, 17.5%,17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%, 19.75%, 20%,20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%,22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%,25.25%, 25.5%, 25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%,27.75%, 28%, 28.25%, 28.5%, 28.75%, 29%, 29.25%, 29.5%, 29.75%, 30%,30.25%, 30.5%, 30.75%, 31%, 31.25%, 31.5%, 31.75%, 32%, 32.25%, 32.5%,32.75%, 33%, 33.25%, 33.5%, 33.75%, 34%, 34.25%, 34.5%, 34.75%, 35%,35.25%, 35.5%, 35.75%, 36%, 36.25%, 36.5%, 36.75%, 37%, 37.25%, 37.5%,37.75%, 38%, 38.25%, 38.5%, 38.75%, 39%, 39.25%, 39.5%, 39.75%, 40%,40.25%, 40.5%, 40.75%, 41%, 41.25%, 41.5%, 41.75%, 42%, 42.25%, 42.5%,42.75%, 43%, 43.25%, 43.5%, 43.75%, 44%, 44.25%, 44.5%, 44.75%, 45%,45.25%, 45.5%, 45.75%, 46%, 46.25%, 46.5%, 46.75%, 47%, 47.25%, 47.5%,47.75%, 48%, 48.25%, 48.5%, 48.75%, 49%, 49.25%, 49.5%, 49.75%, or 50%of the nutritional composition, following centrifugation at 1,000×g for1 hour at 20° C.

Lutein may be in the aqueous phase at about 0.001% to about 30% of thenutritional composition, following centrifugation at 31,000×g for 1 hourat 20° C. Lutein may be in the aqueous phase at about 0.1% to about 20%of the nutritional composition, following centrifugation at 31,000×g for1 hour at 20° C. Lutein may be in the aqueous phase at about 1% to about5% of the nutritional composition, following centrifugation at 31,000×gfor 1 hour at 20° C. For example, lutein may be in the assembly at about0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%,0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.25%,1.5%, 1.75%, 2%, 2.25%, 2.5%, 2.75%, 3%, 3.25%, 3.5%, 3.75%, 4%, 4.25%,4.5%, 4.75%, 5%, 5.25%, 5.5%, 5.75%, 6%, 6.25%, 6.5%, 6.75%, 7%, 7.25%,7.5%, 7.75%, 8%, 8.25%, 8.5%, 8.75%, 9%, 9.25%, 9.5%, 9.75%, 10%,10.25%, 10.5%, 10.75%, 11%, 11.25%, 11.5%, 11.75%, 12%, 12.25%, 12.5%,12.75%, 13%, 13.25%, 13.5%, 13.75%, 14%, 14.25%, 14.5%, 14.75%, 15%,15.25%, 15.5%, 15.75%, 16%, 16.25%, 16.5%, 16.75%, 17%, 17.25%, 17.5%,17.75%, 18%, 18.25%, 18.5%, 18.75%, 19%, 19.25%, 19.5%, 19.75%, 20%,20.25%, 20.5%, 20.75%, 21%, 21.25%, 21.5%, 21.75%, 22%, 22.25%, 22.5%,22.75%, 23%, 23.25%, 23.5%, 23.75%, 24%, 24.25%, 24.5%, 24.75%, 25%,25.25%, 25.5%, 25.75%, 26%, 26.25%, 26.5%, 26.75%, 27%, 27.25%, 27.5%,27.75%, 28%, 28.25%, 28.5%, 28.75%, 29%, 29.25%, 29.5%, 29.75%, or 30%of the nutritional composition, following centrifugation at 31,000×g for1 hour at 20° C.

Lutein may be in the aqueous phase at about 0.001% to about 20% of thenutritional composition, following centrifugation at 100,000×g for 1hour at 20° C. Lutein may be in the aqueous phase at about 0.01% toabout 15% of the nutritional composition, following centrifugation at100,000×g for 1 hour at 20° C. Lutein may be in the aqueous phase atabout 0.1% to about 10% of the nutritional composition, followingcentrifugation at 100,000×g for 1 hour at 20° C. Lutein may be in theaqueous phase at about 1% to about 5% of the nutritional composition,following centrifugation at 100,000×g for 1 hour at 20° C. For example,lutein may be in the aqueous phase at about 0.001%, 0.05%, 0.1%, 0.15%,0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%,0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%,1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%,2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%,4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%,5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%,6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%,7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%,8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%,10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%,11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12%,12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%, 12.7%, 12.8%, 12.9%, 13%,13.1%, 13.2%, 13.3%, 13.4%, 13.5%, 13.6%, 13.7%, 13.8%, 13.9%, 14%,14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%, 14.7%, 14.8%, 14.9%, 15%,15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%,16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%,17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%,18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%,19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20% ofthe nutritional composition, following centrifugation at 100,000×g for 1hour at 20° C.

Lutein may be in the assembly at about 0.001% to about 20% of thenutritional composition, following centrifugation at 31,000×g for 1 hourat 20° C. Lutein may be in the assembly at about 0.01% to about 15% ofthe nutritional composition, following centrifugation at 31,000×g for 1hour at 20° C. Lutein may be in the assembly at about 0.1% to about 10%of the nutritional composition, following centrifugation at 31,000×g for1 hour at 20° C. Lutein may be in the assembly at about 1% to about 5%of the nutritional composition, following centrifugation at 31,000×g for1 hour at 20° C. For example, lutein may be in the assembly at about0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%,0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%,1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%,2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%,3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%,4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%,6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%,7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%,8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%,9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%,10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%,11.7%, 11.8%, 11.9%, 12%, 12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%,12.7%, 12.8%, 12.9%, 13%, 13.1%, 13.2%, 13.3%, 13.4%, 13.5%, 13.6%,13.7%, 13.8%, 13.9%, 14%, 14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%,14.7%, 14.8%, 14.9%, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%,15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%,16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%,17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%,18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%,19.7%, 19.8%, 19.9%, or 20% of the nutritional composition, followingcentrifugation at 31,000×g for 1 hour at 20° C.

MDG plays a critical role in the formation and stability of thecompounds associated with the assembly, wherein MDG increases both theconcentration of protein and bound lipophile that remains in the aqueousphase following centrifugation, relative to controls without MDG. MDG inthe nutritional composition may increase the concentration of protein inthe aqueous phase by about 0.001% to about 20%, following centrifugationat 31,000×g for 1 hour at 20° C., compared to controls without MDG. MDGin the nutritional composition may increase the concentration of proteinin the aqueous phase by about 0.01% to about 15%, followingcentrifugation at 31,000×g for 1 hour at 20° C., compared to controlswithout MDG. MDG in the nutritional composition may increase theconcentration of protein in the aqueous phase by about 0.1% to about10%, following centrifugation at 31,000×g for 1 hour at 20° C., comparedto controls without MDG. MDG in the nutritional composition may increasethe concentration of protein in the aqueous phase by about 1% to about5%, following centrifugation at 31,000×g for 1 hour at 20° C., comparedto controls without MDG. For example protein may be increased in theaqueous phase by about 0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%,0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%,0.9%, 0.95%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%,2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%,3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%,4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%,5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%,6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%,8%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%,9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%,10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%,11.3%, 11.4%, 11.5%, 11.6%, 11.7%, 11.8%, 11.9%, 12%, 12.1%, 12.2%,12.3%, 12.4%, 12.5%, 12.6%, 12.7%, 12.8%, 12.9%, 13%, 13.1%, 13.2%,13.3%, 13.4%, 13.5%, 13.6%, 13.7%, 13.8%, 13.9%, 14%, 14.1%, 14.2%,14.3%, 14.4%, 14.5%, 14.6%, 14.7%, 14.8%, 14.9%, 15%, 15.1%, 15.2%,15.3%, 15.4%, 15.5%, 15.6%, 15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%,16.3%, 16.4%, 16.5%, 16.6%, 16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%,17.3%, 17.4%, 17.5%, 17.6%, 17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%,18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%,19.3%, 19.4%, 19.5%, 19.6%, 19.7%, 19.8%, 19.9%, or 20%, followingcentrifugation at 31,000×g for 1 hour at 20° C., compared to controlswithout MDG.

MDG in the nutritional composition may increase the bound lipophiliccompound in the aqueous phase by about 0.001% to about 20%, followingcentrifugation at 31,000×g for 1 hour at 20° C., compared to controlswithout MDG. MDG in the nutritional composition may increase the boundlipophilic compound in the aqueous phase by about 0.01% to about 15%,following centrifugation at 31,000×g for 1 hour at 20° C., compared tocontrols without MDG. MDG in the nutritional composition may increasethe bound lipophilic compound in the aqueous phase by about 0.1% toabout 10%, following centrifugation at 31,000×g for 1 hour at 20° C.,compared to controls without MDG. MDG in the nutritional composition mayincrease the bound lipophilic compound in the aqueous phase by about 1%to about 5%, following centrifugation at 31,000×g for 1 hour at 20° C.,compared to controls without MDG. For example, the bound lipophiliccompound may be increased in the aqueous phase and the assembly by about0.001%, 0.05%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%,0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1%, 1.1%,1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%,2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%,3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%,4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%,6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%,7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 8.1%, 8.2%, 8.3%,8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%,9.6%, 9.7%, 9.8%, 9.9%, 10%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%,10.7%, 10.8%, 10.9%, 11%, 11.1%, 11.2%, 11.3%, 11.4%, 11.5%, 11.6%,11.7%, 11.8%, 11.9%, 12%, 12.1%, 12.2%, 12.3%, 12.4%, 12.5%, 12.6%,12.7%, 12.8%, 12.9%, 13%, 13.1%, 13.2%, 13.3%, 13.4%, 13.5%, 13.6%,13.7%, 13.8%, 13.9%, 14%, 14.1%, 14.2%, 14.3%, 14.4%, 14.5%, 14.6%,14.7%, 14.8%, 14.9%, 15%, 15.1%, 15.2%, 15.3%, 15.4%, 15.5%, 15.6%,15.7%, 15.8%, 15.9%, 16%, 16.1%, 16.2%, 16.3%, 16.4%, 16.5%, 16.6%,16.7%, 16.8%, 16.9%, 17%, 17.1%, 17.2%, 17.3%, 17.4%, 17.5%, 17.6%,17.7%, 17.8%, 17.9%, 18%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%,18.7%, 18.8%, 18.9%, 19%, 19.1%, 19.2%, 19.3%, 19.4%, 19.5%, 19.6%,19.7%, 19.8%, 19.9%, or 20%, following centrifugation at 31,000×g for 1hour at 20° C., compared to controls without MDG.

The size of the assembly is dependent on components that make up thenutritional powder, such as the MDG, lipophilic compound(s), protein(s),etc. The size of the assembly may be critical in the bioavailability ofthe lipophilic compound. For example, lutein incorporation wouldincrease with size of the assembly. However, assembly sizes greater thanoutside of the range of the invention, may result in complications withsolubility and digestibility following consumption. The assembly sizemay be measured via size exclusion chromatography techniques know withinthe art and then compared to a known calibration curve. The assemblysize may also be measured by size analysis techniques known within theart that include, but are not limited to, laser diffraction, dynamiclight scattering, sieve separation analysis and image analysis (e.g.,using a microscopic method such as light microscopy or scanning electronmicroscopy). The assembly size may be measured by a Malvern ZetasizerNano NS. In some embodiments, the nutritional composition may becentrifuged as described above, and then the aqueous fraction may beanalyzed by dynamic light scattering machine. Additionally, the size ofthe assembly may be measured via size exclusion chromatography (SEC),following the centrifugation step as described above. This is performedby using reference proteins and their SEC elution times and molecularweight. This then can be used to estimate the size of the assembly. Byextrapolation from a reference protein plot. The size of the assemblymay be about 15 kD to about 3000 kD. The size of the assembly may beabout 100 kD to 1000 kD. The size of the assembly may be about 100 kD toabout 500 kD. The size of the assembly may be about 60 kD to about 400kD. For example, the size of the assembly may be about 15 kD, 20 kD, 25kD, 30 kD, 35 kD, 40 kD, 45 kD, 50 kD, 55 kD, 60 kD, 65 kD, 70 kD, 75kD, 80 kD, 85 kD, 90 kD, 95 kD, 100 kD, 105 kD, 110 kD, 11510, 120 kD,125 kD, 130 kD, 135 kD, 140 kD, 145 kD, 150 kD, 155 kD, 160 kD, 165 kD,170 kD, 175 kD, 180 kD, 185 kD, 190 kD, 195 kD, 200 kD, 205 kD, 210 kD,215 kD, 22010, 225 kD, 230 kD, 235 kD, 240 kD, 245 kD, 250 kD, 255 kD,260 kD, 265 kD, 270 kD, 275 kD, 280 kD, 285 kD, 290 kD, 295 kD, 300 kD,305 kD, 310 kD, 315 kD, 320 kD, 325 kD, 330 kD, 335 kD, 340 kD, 345 kD,350 kD, 355 kD, 360 kD, 365 kD, 370 kD, 375 kD, 380 kD, 385 kD, 390 kD,395 kD, 400 kD, 405 kD, 410 kD, 415 kD, 420 kD, 425 kD, 43010, 435 kD,440 kD, 445 kD, 450 kD, 455 kD, 460 kD, 465 kD, 470 kD, 475 kD, 480 kD,485 kD, 490 kD, 495 kD, 500 kD, 505 kD, 510 kD, 515 kD, 520 kD, 525 kD,530 kD, 535 kD, 540 kD, 545 kD, 550 kD, 555 kD, 560 kD, 565 kD, 570 kD,575 kD, 580 kD, 585 kD, 590 kD, 595 kD, 600 kD, 605 kD, 610 kD, 615 kD,620 kD, 625 kD, 630 kD, 635 kD, 640 kD, 645 kD, 650 kD, 655 kD, 660 kD,665 kD, 670 kD, 675 kD, 680 kD, 685 kD, 690 kD, 695 kD, 700 kD, 705 kD,710 kD, 715 kD, 720 kD, 725 kD, 730 kD, 735 kD, 740 kD, 74510, 750 kD,755 kD, 760 kD, 765 kD, 770 kD, 775 kD, 780 kD, 785 kD, 790 kD, 795 kD,800 kD, 805 kD, 810 kD, 815 kD, 820 kD, 825 kD, 830 kD, 835 kD, 840 kD,845 kD, 85010, 855 kD, 860 kD, 865 kD, 870 kD, 875 kD, 880 kD, 885 kD,890 kD, 895 kD, 900 kD, 905 kD, 910 kD, 915 kD, 920 kD, 925 kD, 930 kD,935 kD, 940 kD, 945 kD, 950 kD, 95510, 960 kD, 965 kD, 970 kD, 975 kD,980 kD, 985 kD, 990 kD, 995 kD, 1000 kD, 1010 kD, 1020 kD, 1030 kD, 1040kD, 1050 kD, 1060 kD, 1070 kD, 1080 kD, 1090 kD, 1100 kD, 1110 kD, 1120kD, 1130 kD, 1140 kD, 1150 kD, 1160 kD, 1170 kD, 1180 kD, 1190 kD, 1200kD, 1210 kD, 1220 kD, 1230 kD, 1240 kD, 1250 kD, 1260 kD, 1270 kD, 1280kD, 1290 kD, 1300 kD, 1310 kD, 1320 kD, 1330 kD, 1340 kD, 1350 kD, 1360kD, 1370 kD, 1380 kD, 1390 kD, 1400 kD, 1410 kD, 1420 kD, 1430 kD, 1440kD, 1450 kD, 1460 kD, 1470 kD, 1480 kD, 1490 kD, 1500 kD, 1510 kD, 1520kD, 1530 kD, 1540 kD, 1550 kD, 1560 kD, 1570 kD, 1580 kD, 1590 kD, 1600kD, 1610 kD, 1620 kD, 1630 kD, 1640 kD, 1650 kD, 1660 kD, 1670 kD, 1680kD, 1690 kD, 1700 kD, 1710 kD, 1720 kD, 1730 kD, 1740 kD, 1750 kD, 1760kD, 1770 kD, 1780 kD, 1790 kD, 1800 kD, 1810 kD, 1820 kD, 1830 kD, 1840kD, 1850 kD, 1860 kD, 1870 kD, 1880 kD, 1890 kD, 1900 kD, 1910 kD, 1920kD, 1930 kD, 1940 kD, 1950 kD, 1960 kD, 1970 kD, 1980 kD, 1990 kD, 2000kD, 2010 kD, 2020 kD, 2030 kD, 2040 kD, 2050 kD, 2060 kD, 2070 kD, 2080kD, 2090 kD, 2100 kD, 2110 kD, 2120 kD, 2130 kD, 2140 kD, 2150 kD, 2160kD, 2170 kD, 2180 kD, 2190 kD, 2200 kD, 2210 kD, 2220 kD, 2230 kD, 2240kD, 2250 kD, 2260 kD, 2270 kD, 2280 kD, 2290 kD, 2300 kD, 2310 kD, 2320kD, 2330 kD, 2340 kD, 2350 kD, 2360 kD, 2370 kD, 2380 kD, 2390 kD, 2400kD, 2410 kD, 2420 kD, 2430 kD, 2440 kD, 2450 kD, 2460 kD, 2470 kD, 2480kD, 2490 kD, 2500 kD, 2510 kD, 2520 kD, 2530 kD, 2540 kD, 2550 kD, 2560kD, 2570 kD, 2580 kD, 2590 kD, 2600 kD, 2610 kD, 2620 kD, 2630 kD, 2640kD, 2650 kD, 2660 kD, 2670 kD, 2680 kD, 2690 kD, 2700 kD, 2710 kD, 2720kD, 2730 kD, 2740 kD, 2750 kD, 2760 kD, 2770 kD, 2780 kD, 2790 kD, 2800kD, 2810 kD, 2820 kD, 2830 kD, 2840 kD, 2850 kD, 2860 kD, 2870 kD, 2880kD, 2890 kD, 2900 kD, 2910 kD, 2920 kD, 2930 kD, 2940 kD, 2950 kD, 2960kD, 2970 kD, 2980 kD, 2990 kD, or 3000 kD.

The nutritional composition may include an assembly that increases thebioavailability of the lipophilic compound from about 5% to about 80%,relative to controls without MDG and hydrophobic protein. Thenutritional composition may include an assembly that increases the rangeof bioavailability of the lipophilic compound from about 40% to about55%, relative to controls without MDG and hydrophobic protein. Thenutritional composition may include an assembly that increases the rangebioavailability of the lipophilic compound from about 70% to about 75%,relative to controls without MDG and hydrophobic protein. Thebioavailability of the lipophilic compound is dependent on the formationof the assemblies of the nutritional composition, wherein MDG andhydrophobic protein are comprised within the assembly. Thebioavailability of the assembly may be assessed via lymphatic analysisin rats. Specifically, rats may be fasted overnight prior to surgery,and under anesthesia, a laparotomy may be performed, and the intestinallymph duct may be cannulated according to the procedure of Tso et al.,“The Absorption of Lipid and Lipoprotein Synthesis,” Lipid ResearchMethodology, Chapter 5: 191-216 (1984) Alan R. Liss, Inc., NY, N.Y.,hereby incorporated by reference to the extent consistent herewith. Thesuperior mesenteric artery is isolated, but not occluded. A siliconinfusion tube (1.6 mm OD) is placed in the stomach for future infusionof a test sample. The fundic incision is closed by a purse stringsuture. Rats are allowed to recover for 24 hours before infusion begins.Animals are intragastrically infused 24 hours after surgery with 3 ml ofnutritional composition that delivers lipophilic compound. Lymph arecollected in pre-cooled tubes 1 hour before the nutritional compositioninfusions (fasting) and then hourly for 8 hours after initiation ofinfusion. At the end of the 8 hour infusion, rats are sacrificed byexsanguination.

The lymph lipids were extracted and analyzed for lutein concentrationusing high-performance liquid chromatography with programmed wavelengthultraviolet detection (Craft Technologies, Wilson, N.C.). For example,the assembly may increase the bioavailability of the lipophilic compoundby about 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%,11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%,17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, 20.5%, 21%, 21.5%, 22%, 22.5%,23%, 23.5%, 24%, 24.5%, 25%, 25.5%, 26%, 26.5%, 27%, 27.5%, 28%, 28.5%,29%, 29.5%, 30%, 30.5%, 31%, 31.5%, 32%, 32.5%, 33%, 33.5%, 34%, 34.5%,35%, 35.5%, 36%, 36.5%, 37%, 37.5%, 38%, 38.5%, 39%, 39.5%, 40%, 40.5%,41%, 41.5%, 42%, 42.5%, 43%, 43.5%, 44%, 44.5%, 45%, 45.5%, 46%, 46.5%,47%, 47.5%, 48%, 48.5%, 49%, 49.5%, 50%, 50.5%, 51%, 51.5%, 52%, 52.5%,53%, 53.5%, 54%, 54.5%, 55%, 55.5%, 56%, 56.5%, 57%, 57.5%, 58%, 58.5%,59%, 59.5%, 60%, 60.5%, 61%, 61.5%, 62%, 62.5%, 63%, 63.5%, 64%, 64.5%,65%, 65.5%, 66%, 66.5%, 67%, 67.5%, 68%, 68.5%, 69%, 69.5%, 70%, 70.5%,71%, 71.5%, 72%, 72.5%, 73%, 73.5%, 74%, 74.5%, 75%, 75.5%, 76%, 76.5%,77%, 77.5%, 78%, 78.5%, 79%, 79.5%, or 80%.

(1) Activated Assembly

The assembly may be an activated assembly. The activated assembly may becomprised in a powder, suspension, solution, emulsion or combinationsthereof. The activated assembly may comprise MDG, at least onelipophilic compound, and at least one hydrophobic protein. The activatedassembly may be in, but is not limited to, a premix, protected premix,and an activated premix. The activated assembly may contact an aqueousliquid as part of the activated premix.

(2) Premix/Protected Premix

The premix may or may not comprise the assembly. The protected premixmay or may not comprise the assembly.

(3) Activated Premix Assembly

The activated premix may comprise the assembly. The assembly may be in aprotected activated premix. The assembly in the activated premix maycomprise MDG, at least one lipophilic compound, and at least onehydrophobic protein. The assembly in the protected activated premix maycomprise MDG, at least one lipophilic compound, and at least onehydrophobic protein. The activated premix comprising the assembly may beadded to additional components of the nutritional composition. Theprotected activated premix comprising the assembly may be added toadditional components of the nutritional composition.

In some embodiments, the premix is contacted to a protein-in-waterslurry, to form an activated premix. This is then added to theadditional components of the nutritional composition. In someembodiments, the premix comprising the assembly is contacted to aprotein-in-oil slurry, to form an activated premix. This is then addedto the additional components of the nutritional composition. In someembodiments, the premix comprising the assembly is contacted to acarbohydrate-mineral slurry and a protein-in-oil slurry, to form anactivated premix. This is then added to the additional components of thenutritional composition. In some embodiments, the premix comprising theassembly is contacted to an oil slurry, to form an activated premix.This is then added to the additional components of the nutritionalcomposition. In some embodiments, the premix comprising the assembly iscontacted to final blend slurry, to form an activated premix. This isthen added to the additional components of the nutritional composition.

(4) Shelf-Life

The nutritional composition may include an assembly with an enhancedshelf-life. The assembly may have an enhanced shelf-life in a, but isnot limited to, powder, premix, activated premix, protected activatedpremix, suspension, liquid, emulsion mixture, or combination thereof.

The nutritional composition may include an assembly that is stable at23° C. for at least about 1 day to at least about 36 months. Thenutritional composition may include an assembly that is stable at 23° C.for at least about 10 days to at least about 36 months. The nutritionalcomposition may include an assembly that is stable at 23° C. for atleast about 6 months to at least about 12 months. The nutritionalcomposition may include an assembly that is stable at 23° C. for atleast about 1 day to at least about 24 months. The stability of theassembly is dependent on the combination of MDG and the hydrophobicprotein's ability to interact with the lipophilic compound, which keepsthe lipophilic compound in solution, thereby enhancing itsbiocompatibility. The stability of the assembly may be investigated bysize exclusion chromatography techniques known within the art atvariable time points post formulating the nutritional composition. Theassembly may be stable at 23° C. for at least about 1 day, 2 days, 3days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days,12 days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days,20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days,28 days, 29 days, 30 days, 31 days, 2 months, 3 months, 4 months, 5months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12months, 13 months, 14 months, 15 months, 16 months, 17 months, 18months, 19 months, 20 months, 21 months, 22 months, 23 months, 24months, 25 months, 26 months, 27 months, 28 months, 29 months, 30months, 31 months, 32 months, 33 months, 34 months, 35 months, or 36months.

c. Lipophilic Compound

The nutritional composition may comprise at least one lipophiliccompound. The lipophilic compound may be present within the premix,activated premix, and/or the protected premix. The lipophilic compoundmay be incorporated into the assembly. Lipophilic compounds have pooraqueous solubility, resulting in decreased bioavailability whenincorporated within a nutritional composition. However, by incorporatingthe lipophilic compound within the aforementioned assembly, along withMDG and the at least one hydrophobic protein, it is more soluble andstable. The lipophilic compound may be a carotenoid, zeaxanthin, alpha-or beta-cryptoxanthin, retinol, a ganglioside, a phosphoinositide, alipoprotein, a phospholipid, a glycolipid, a glycophospholipid, an oilsoluble vitamin such as synthetic and natural forms of vitamins A, E, D,or K, lycopene or mixtures thereof. The lipophilic compound may be asource of long chain polyunsaturated fatty acids (LCPUFAs). Someexamples of LCPUFAs include, but are not limited, to eicosapentaenoicacid (“EPA”), arachidonic acid (ARA), linoleic (18:2 n-6), γ-linolenic(18:3 n-6), dihomo-γ-linolenic (20:3 n-6) acids in the n-6 pathway,α-linolenic (18:3 n-3), stearidonic (18:4 n-3), eicosatetraenoic (20:4n-3), eicosapentaenoic (20:5 n-3), and DHA (22:6 n-3). The lipophiliccompound may be lutein. Additionally, the lipophilic compound may be amixture of different lipophilic compounds.

Suitable, non-limiting examples of lipophilic pharmaceutical agents thatmay be included in the present composition may be selected from: oilsoluble drugs (e.g., immunosuppressive agents such as Cyclosporin™,protease inhibitors such as Ritonavir™ macrolide antibiotics and oilsoluble anesthetics such as Propofol™); synthetic and natural forms ofsteroidal hormones such as estrogens, estradiols, progesterone,testosterone, cortisone, phytoestrogens, dehydroepiandrosterone (DHEA),and growth hormones; oil soluble acids and alcohols (e.g., tartaricacid, lactylic acid, butylated hydroxyanisole, butylated hydroxytoluene,lignin, phytosterols, flavonoids such as quercetin and resveratrol, anddiallyl disulfides); and combinations thereof. Additional lipophilicpharmaceutical agents may be found in U.S. patent application Ser. No.13/452,033, which was filed on Apr. 20, 2012 and is hereby incorporatedby reference to the extent that it is consistent with the presentdisclosure.

The lipophilic compound is a valuable source of nutrients for infants,toddlers, children and adults. The lipophilic compound may be used fortreating or maintaining appropriate health, such as, but is not limitedto, bone, eye, cardiovascular, and cerebral health.

(1) Lutein

The lipophilic compound of the nutritional composition may be lutein.Lutein may be incorporated into the assembly. Incorporation of lutein aspart of the assembly enhances its solubility, thus enhancing itsbioavailability for treatment of health conditions, relative to controlswithout MDG or hydrophobic protein. The term “lutein” as used herein,unless otherwise specified, refers to one or more of free lutein, luteinesters, lutein salts, or other lutein derivatives or related structuresas described or otherwise suggested herein. Lutein or lutein sourcessuitable for use in the nutritional composition of the presentdisclosure include free lutein as well as esters, salts or otherderivatives or related structures thereof, including those that conformto the formula:

The above formula includes the general structure of lutein and relatedderivatives or structures. Free lutein, for example, corresponds to theformula wherein R₁ and R₂ are both hydrogen, and includes cis and transisomers thereof as well as salts thereof, e.g., sodium, potassium.

Lutein esters suitable for use in the nutritional composition of thepresent disclosure include any lutein ester of the above formula whereinR₁ and R₂ are the same or different, and are nutritionally acceptablemonovalent salts, hydrogen or an acyl residue of a carboxylic acid,provided that at least one of R₁ and R₂ is an acyl residue of acarboxylic acid. Suitable lutein esters include, as well, both cis andtrans isomers. The R₁ and R₂ moieties are residues of saturated orunsaturated C₁ to C₂₂ fatty carboxylic acids, non-limiting examples ofwhich include formic, acetic, propionic, butyric, valeric, caproic,caprylic, capric, lauric, myristic, palmitic, stearic, and oleic acids.

Any natural or synthetic source of lutein is suitable for use hereinprovided that such a source is also known for or otherwise suitable foruse in nutritional composition and is compatible with the other selectedingredients within the composition. Lutein sources may be provided asindividual ingredients or in any combination with other materials orsources, including sources such as multivitamin premixes, mixedcarotenoid premixes, and pure lutein sources. In some embodiments,components of the nutritional composition may inherently comprise lutein(e.g., lutein rich oils, surfactants, etc.).

Non-limiting examples of suitable lutein sources for use herein includeFloraGLO® Crystalline Lutein, available from Kemin Foods, Des Moines,Iowa, USA; and LUTIMAX Lutein Esters provided by OmniActive, Mumbai,India.

In some embodiments, it may be desirable to blend lutein with the MDGoil to form an MDG premix before blending the MDG oil or lutein withother ingredients of the composition. To make the MDG premix, lutein maybe mixed with the MDG oil in the presence of heat or at ambienttemperature and, in some embodiments, with agitation to allow the luteinto dissolve, disperse, or suspend into the MDG oil.

In some embodiments, it may be desirable to form an MDG-protected premixcomprising lutein, a surfactant, and an MDG oil before blending the MDGoil or lutein with other ingredients of the composition. In theseembodiments, lutein and a surfactant may be mixed with the MDG oil inthe presence of heat or at ambient temperature and, in some embodiments,with agitation to allow the lutein to dissolve, disperse, or suspend inthe MDG oil.

Lutein is a good source of nutrient for infants, toddlers, children andadults. Lutein may be used within the nutritional composition tomaintain and supplement healthy vision and eye care. Lutein may be usedwithin the nutritional composition to maintain and supplement cerebralhealth and development.

Lutein may in the nutritional composition at about 0.01 mg/kg to about50 mg/kg. Lutein may in the nutritional composition at about 0.02 mg/kgto about 20 mg/kg. Lutein may in the nutritional composition at about0.1 mg/kg to about 20 mg/kg. Lutein may in the nutritional compositionat about 1 mg/kg to about 15 mg/kg. For example, lutein may be in thenutritional composition at about 0.01 mg/kg, 0.02 mg/kg, 0.03 mg/kg,0.04 mg/kg, 0.05 mg/kg, 0.1 mg/kg, 0.25 mg/kg, 0.5 mg/kg, 0.75 mg/kg, 1mg/kg, 1.25 mg/kg, 1.5 mg/kg, 1.75 mg/kg, 2 mg/kg, 2.25 mg/kg, 2.5mg/kg, 2.75 mg/kg, 3 mg/kg, 3.25 mg/kg, 3.5 mg/kg, 3.75 mg/kg, 4 mg/kg,4.25 mg/kg, 4.5 mg/kg, 4.75 mg/kg, 5 mg/kg, 5.25 mg/kg, 5.5 mg/kg, 5.75mg/kg, 6 mg/kg, 6.25 mg/kg, 6.5 mg/kg, 6.75 mg/kg, 7 mg/kg, 7.25 mg/kg,7.5 mg/kg, 7.75 mg/kg, 8 mg/kg, 8.25 mg/kg, 8.5 mg/kg, 8.75 mg/kg, 9mg/kg, 9.25 mg/kg, 9.5 mg/kg, 9.75 mg/kg, 10 mg/kg, 10.25 mg/kg, 10.5mg/kg, 10.75 mg/kg, 11 mg/kg, 11.25 mg/kg, 11.5 mg/kg, 11.75 mg/kg, 12mg/kg, 12.25 mg/kg, 12.5 mg/kg, 12.75 mg/kg, 13 mg/kg, 13.25 mg/kg, 13.5mg/kg, 13.75 mg/kg, 14 mg/kg, 14.25 mg/kg, 14.5 mg/kg, 14.75 mg/kg, 15mg/kg, 15.25 mg/kg, 15.5 mg/kg, 15.75 mg/kg, 16 mg/kg, 16.25 mg/kg, 16.5mg/kg, 16.75 mg/kg, 17 mg/kg, 17.25 mg/kg, 17.5 mg/kg, 17.75 mg/kg, 18mg/kg, 18.25 mg/kg, 18.5 mg/kg, 18.75 mg/kg, 19 mg/kg, 19.25 mg/kg, 19.5mg/kg, 19.75 mg/kg, 20 mg/kg, 20.25 mg/kg, 20.5 mg/kg, 20.75 mg/kg, 21mg/kg, 21.25 mg/kg, 21.5 mg/kg, 21.75 mg/kg, 22 mg/kg, 22.25 mg/kg, 22.5mg/kg, 22.75 mg/kg, 23 mg/kg, 23.25 mg/kg, 23.5 mg/kg, 23.75 mg/kg, 24mg/kg, 24.25 mg/kg, 24.5 mg/kg, 24.75 mg/kg, 25 mg/kg, 25.25 mg/kg, 25.5mg/kg, 25.75 mg/kg, 26 mg/kg, 26.25 mg/kg, 26.5 mg/kg, 26.75 mg/kg, 27mg/kg, 27.25 mg/kg, 27.5 mg/kg, 27.75 mg/kg, 28 mg/kg, 28.25 mg/kg, 28.5mg/kg, 28.75 mg/kg, 29 mg/kg, 29.25 mg/kg, 29.5 mg/kg, 29.75 mg/kg, 30mg/kg, 30.25 mg/kg, 30.5 mg/kg, 30.75 mg/kg, 31 mg/kg, 31.25 mg/kg, 31.5mg/kg, 31.75 mg/kg, 32 mg/kg, 32.25 mg/kg, 32.5 mg/kg, 32.75 mg/kg, 33mg/kg, 33.25 mg/kg, 33.5 mg/kg, 33.75 mg/kg, 34 mg/kg, 34.25 mg/kg, 34.5mg/kg, 34.75 mg/kg, 35 mg/kg, 35.25 mg/kg, 35.5 mg/kg, 35.75 mg/kg, 36mg/kg, 36.25 mg/kg, 36.5 mg/kg, 36.75 mg/kg, 37 mg/kg, 37.25 mg/kg, 37.5mg/kg, 37.75 mg/kg, 38 mg/kg, 38.25 mg/kg, 38.5 mg/kg, 38.75 mg/kg, 39mg/kg, 39.25 mg/kg, 39.5 mg/kg, 39.75 mg/kg, 40 mg/kg, 40.25 mg/kg, 40.5mg/kg, 40.75 mg/kg, 41 mg/kg, 41.25 mg/kg, 41.5 mg/kg, 41.75 mg/kg, 42mg/kg, 42.25 mg/kg, 42.5 mg/kg, 42.75 mg/kg, 43 mg/kg, 43.25 mg/kg, 43.5mg/kg, 43.75 mg/kg, 44 mg/kg, 44.25 mg/kg, 44.5 mg/kg, 44.75 mg/kg, 45mg/kg, 45.25 mg/kg, 45.5 mg/kg, 45.75 mg/kg, 46 mg/kg, 46.25 mg/kg, 46.5mg/kg, 46.75 mg/kg, 47 mg/kg, 47.25 mg/kg, 47.5 mg/kg, 47.75 mg/kg, 48mg/kg, 48.25 mg/kg, 48.5 mg/kg, 48.75 mg/kg, 49 mg/kg, 49.25 mg/kg, 49.5mg/kg, 49.75 mg/kg, or 50 mg/kg.

(2) Vitamin D

The lipophilic compound of the nutritional composition may be vitamin D.Vitamin D may be incorporated into the assembly. Incorporation ofvitamin D as part of the assembly enhances its solubility, thusenhancing its bioavailability for treatment of health conditions,relative to controls without MDG or hydrophobic protein. “Vitamin D”refers to a group of lipophilic compounds, or “vitamers,” related tosteroids. There are several vitamers encompassed by the term “VitaminD,” but the most important of these are ergocalciferol, also known asVitamin D₂:

and cholecalciferol, also known as Vitamin D₃:

For the purpose of this disclosure, the term “Vitamin D” as used herein,unless otherwise specified, encompasses all forms of Vitamin D, whetheras individual vitamers such as Vitamin D2 or Vitamin D3, or ascombinations of two or more vitamers.

Vitamin D may be ingested from the diet, and Vitamin D3 is alsosynthesized in mammalian skin by the reaction of cholesterol with UVradiation from sunlight. Once in the body, Vitamin D vitamers aremetabolized into other chemical forms that regulate the concentration ofcalcium and phosphate in the bloodstream and promote the healthy growthand maintenance of bone. Vitamin D may be used within the nutritionalcomposition to maintain and supplement healthy bone care andmaintenance. Vitamin D may be used within the nutritional compositionfor functions to activate the innate and dampen the adaptive immunesystems and assist in cognitive development.

Any natural or synthetic source of Vitamin D is suitable for use in thecomposition disclosed herein, provided that such a source is suitablefor use in ingestible the composition and is compatible with the otheringredients in the composition. Vitamin D may be provided as anindividual ingredient or in any combination with other materials orsources, including sources such as multivitamin premixes. For example,Vitamin D may be mixed with other oil-soluble vitamins such as VitaminA, Vitamin E, or Vitamin K.

Non-limiting examples of suitable Vitamin D sources for use hereininclude Vitamin D3 provided by BASF Corporation (Florham Park, N.J.,USA), Vitamin D3 provided by Fermenta Biotech Ltd. (Kullu, HimachalPradesh, India), or Quali®-D provided by DSM Nutritional Products AG(Kaiseraugst, Switzerland).

Vitamin D may be in the nutritional composition at about 1 μg/kg toabout 30 μg/kg. Vitamin D may be in the nutritional composition at about10 μg/kg to about 30 μg/kg. Vitamin D may be in the nutritionalcomposition at about 5 μg/kg to about 25 μg/kg. Vitamin D may be in thenutritional composition at about 10 μg/kg to about 20 μg/kg. Forexample, vitamin D may be in the nutritional composition at about 1μg/kg, 1.2 μg/kg, 1.4 μg/kg, 1.6 μg/kg, 1.8 μg/kg, 2 μg/kg, 2.2 μg/kg,2.4 μg/kg, 2.6 μg/kg, 2.8 μg/kg, 3 μg/kg, 3.2 μg/kg, 3.4 μg/kg, 3.6μg/kg, 3.8 μg/kg, 4 μg/kg, 4.2 μg/kg, 4.4 μg/kg, 4.6 μg/kg, 4.8 μg/kg, 5μg/kg, 5.2 μg/kg, 5.4 μg/kg, 5.6 μg/kg, 5.8 μg/kg, 6 μg/kg, 6.2 μg/kg,6.4 μg/kg, 6.6 μg/kg, 6.8 μg/kg, 7 μg/kg, 7.2 μg/kg, 7.4 μg/kg, 7.6μg/kg, 7.8 μg/kg, 8 μg/kg, 8.2 μg/kg, 8.4 μg/kg, 8.6 μg/kg, 8.8 μg/kg, 9μg/kg, 9.2 μg/kg, 9.4 μg/kg, 9.6 μg/kg, 9.8 μg/kg, 10 μg/kg, 10.2 μg/kg,10.4 μg/kg, 10.6 μg/kg, 10.8 μg/kg, 11 μg/kg, 11.2 μg/kg, 11.4 μg/kg,11.6 μg/kg, 11.8 μg/kg, 12 μg/kg, 12.2 μg/kg, 12.4 μg/kg, 12.6 μg/kg,12.8 μg/kg, 13 μg/kg, 13.2 μg/kg, 13.4 μg/kg, 13.6 μg/kg, 13.8 μg/kg, 14μg/kg, 14.2 μg/kg, 14.4 μg/kg, 14.6 μg/kg, 14.8 μg/kg, 15 μg/kg, 15.2μg/kg, 15.4 μg/kg, 15.6 μg/kg, 15.8 μg/kg, 16 μg/kg, 16.2 μg/kg, 16.4μg/kg, 16.6 μg/kg, 16.8 μg/kg, 17 μg/kg, 17.2 μg/kg, 17.4 μg/kg, 17.6μg/kg, 17.8 μg/kg, 18 μg/kg, 18.2 μg/kg, 18.4 μg/kg, 18.6 μg/kg, 18.8μg/kg, 19 μg/kg, 19.2 μg/kg, 19.4 μg/kg, 19.6 μg/kg, 19.8 μg/kg, 20μg/kg, 20.2 μg/kg, 20.4 μg/kg, 20.6 μg/kg, 20.8 μg/kg, 21 μg/kg, 21.2μg/kg, 21.4 μg/kg, 21.6 μg/kg, 21.8 μg/kg, 22 μg/kg, 22.2 μg/kg, 22.4μg/kg, 22.6 μg/kg, 22.8 μg/kg, 23 μg/kg, 23.2 μg/kg, 23.4 μg/kg, 23.6μg/kg, 23.8 μg/kg, 24 μg/kg, 24.2 μg/kg, 24.4 μg/kg, 24.6 μg/kg, 24.8μg/kg, 25 μg/kg, 25.2 μg/kg, 25.4 μg/kg, 25.6 μg/kg, 25.8 μg/kg, 26μg/kg, 26.2 μg/kg, 26.4 μg/kg, 26.6 μg/kg, 26.8 μg/kg, 27 μg/kg, 27.2μg/kg, 27.4 μg/kg, 27.6 μg/kg, 27.8 μg/kg, 28 μg/kg, 28.2 μg/kg, 28.4μg/kg, 28.6 μg/kg, 28.8 μg/kg, 29 μg/kg, 29.2 μg/kg, 29.4 μg/kg, 29.6μg/kg, 29.8 μg/kg, or 30 μg/kg.

(3) Vitamin E

The lipophilic compound of the nutritional composition may be vitamin E.Vitamin E may be incorporated into the assembly. Incorporation ofvitamin E as part of the assembly enhances its solubility, thusenhancing its bioavailability for treatment of health conditions,relative to controls without MDG or hydrophobic protein. Vitamin E maybe used within the nutritional composition to provide antioxidants andmaintain and supplement cardiovascular health. Vitamin E may also bereferred to as tocopherol. Tocopherols are available in four forms,alpha, beta, gamma, and delta, which differ in the number and positionof the methyl groups on the chroman ring (see the table 1 below).Further, tocopherols may exist in a number of stereoisomeric formsdepending on the chirality of the phytyl tail. Of the alpha tocopherols,RRR-alpha tocopherol (also referred to as “natural vitamin E”) has thegreatest biological activity and is reported to be the dominant form ofthe alpha tocopherol in the brain. In one aspect, the composition maycomprise RRR-alpha tocopherol, which means that the composition iseither formulated with the addition of RRR-alpha tocopherol or areotherwise prepared so as to contain RRR-alpha tocopherol. As usedherein, the term “RRR-alpha tocopherol” refers to both exogenous sourcesand inherent sources of free RRR-alpha tocopherol and RRR-alphatocopherol esters such as RRR alpha tocopherol acetate that are presentin a composition. Inherent sources include RRR-alpha tocopherol that isinherently present in components that are present in a nutritionalcomposition and may include for example, various oils and fats.Exogenous sources of RRR-alpha tocopherol include RRR-alpha tocopherolacetate that is added to the nutritional composition not as part ofanother component. Any source of RRR-alpha tocopherol is suitable foruse herein provided that the finished product contains RRR-alphatocopherol.

RRR-alpha tocopherol is a single stereoisomer whereas synthetic vitaminE (all-rac-alpha tocopherol or tocopherol acetate) is an equimolarmixture of eight isomers, only one of which is RRR-alpha tocopherol. Thefact that the dominant form of alpha tocopherol is RRR alpha tocopherol(based on animal studies) strongly suggests that the other seven chiralisomers must be absorbed at a lower rate by the brain or oxidized at afaster rate. Cholesterol is a major component of myelin, and it islikely that stimulated cholesterol synthesis may stimulate newborninfant neuron myelination.

TABLE 1 Types of Tocopherols Compound R₁ R₂ R₃ alpha-tocopherol Me Me Mebeta-tocopherol Me H Me gamma-tocopherol H Me Me delta-tocopherol H H Me

Vitamin E may be in the nutritional composition at about 5 mg/kg toabout 75 mg/kg. Vitamin E may be in the nutritional composition at about10 mg/kg to about 60 mg/kg. Vitamin E may be in the nutritionalcomposition at about 20 mg/kg to about 40 mg/kg. For example, vitamin Emay be in the nutritional composition at about 5 mg/kg, 5.5 mg/kg, 6mg/kg, 6.5 mg/kg, 7 mg/kg, 7.5 mg/kg, 8 mg/kg, 8.5 mg/kg, 9 mg/kg, 9.5mg/kg, 10 mg/kg, 10.5 mg/kg, 11 mg/kg, 11.5 mg/kg, 12 mg/kg, 12.5 mg/kg,13 mg/kg, 13.5 mg/kg, 14 mg/kg, 14.5 mg/kg, 15 mg/kg, 15.5 mg/kg, 16mg/kg, 16.5 mg/kg, 17 mg/kg, 17.5 mg/kg, 18 mg/kg, 18.5 mg/kg, 19 mg/kg,19.5 mg/kg, 20 mg/kg, 20.5 mg/kg, 21 mg/kg, 21.5 mg/kg, 22 mg/kg, 22.5mg/kg, 23 mg/kg, 23.5 mg/kg, 24 mg/kg, 24.5 mg/kg, 25 mg/kg, 25.5 mg/kg,26 mg/kg, 26.5 mg/kg, 27 mg/kg, 27.5 mg/kg, 28 mg/kg, 28.5 mg/kg, 29mg/kg, 29.5 mg/kg, 30 mg/kg, 30.5 mg/kg, 31 mg/kg, 31.5 mg/kg, 32 mg/kg,32.5 mg/kg, 33 mg/kg, 33.5 mg/kg, 34 mg/kg, 34.5 mg/kg, 35 mg/kg, 35.5mg/kg, 36 mg/kg, 36.5 mg/kg, 37 mg/kg, 37.5 mg/kg, 38 mg/kg, 38.5 mg/kg,39 mg/kg, 39.5 mg/kg, 40 mg/kg, 40.5 mg/kg, 41 mg/kg, 41.5 mg/kg, 42mg/kg, 42.5 mg/kg, 43 mg/kg, 43.5 mg/kg, 44 mg/kg, 44.5 mg/kg, 45 mg/kg,45.5 mg/kg, 46 mg/kg, 46.5 mg/kg, 47 mg/kg, 47.5 mg/kg, 48 mg/kg, 48.5mg/kg, 49 mg/kg, 49.5 mg/kg, 50 mg/kg, 50.5 mg/kg, 51 mg/kg, 51.5 mg/kg,52 mg/kg, 52.5 mg/kg, 53 mg/kg, 53.5 mg/kg, 54 mg/kg, 54.5 mg/kg, 55mg/kg, 55.5 mg/kg, 56 mg/kg, 56.5 mg/kg, 57 mg/kg, 57.5 mg/kg, 58 mg/kg,58.5 mg/kg, 59 mg/kg, 59.5 mg/kg, 60 mg/kg, 60.5 mg/kg, 61 mg/kg, 61.5mg/kg, 62 mg/kg, 62.5 mg/kg, 63 mg/kg, 63.5 mg/kg, 64 mg/kg, 64.5 mg/kg,65 mg/kg, 65.5 mg/kg, 66 mg/kg, 66.5 mg/kg, 67 mg/kg, 67.5 mg/kg, 68mg/kg, 68.5 mg/kg, 69 mg/kg, 69.5 mg/kg, 70 mg/kg, 70.5 mg/kg, 71 mg/kg,71.5 mg/kg, 72 mg/kg, 72.5 mg/kg, 73 mg/kg, 73.5 mg/kg, 74 mg/kg, 74.5mg/kg, or 75 mg/kg.

(4) Docosahexaenoic Acid (DHA)

The lipophilic compound of the nutritional composition may be DHA. DHAmay be incorporated into the assembly. Incorporation of DHA as part ofthe assembly enhances its solubility, thus enhancing its bioavailabilityfor treatment of health conditions, relative to controls without MDG orhydrophobic protein. DHA may be used within the nutritional compositionto maintain and supplement conditions such as healthy brain function.DHA is an omega-3 fatty acid with 22 carbons in the lipophilic chain,typically found, for example, in cold water fish and algae:

DHA is a primary structural component of the human brain, cerebralcortex, and retina, and is an essential fatty acid for properdevelopment of the eyes, brain, and nervous system of infants andbabies. Breast milk contains DHA, and many infant formulas are enrichedwith DHA because of its crucial role in the growth and development ofthese systems in infants. There is increasing interest in providingsupplemental DHA to pregnant women. DHA is believed to lower the risk ofdeveloping lacrimal keratoconjunctivitis, glaucoma, and maculardegeneration. DHA is also needed for healthy brain functioning inadults. DHA deficiency may contribute to the decline in mentalfunctioning of healthy older adults, and preliminary studies indicatethat early intervention with DHA supplements may improve memory andlearning in adults over 50 years of age.

Non-limiting examples of suitable DHA sources for use herein includeLife's DHA®, available from DSM Nutritional Products, Kaiseraugst,Switzerland, and Maxomega™ DHA, available from BASF Pharma Ltd.,Callanish, Scotland.

DHA may be in the nutritional composition at about 10 mg/kg to about 10g/kg. DHA may be in the nutritional composition at about 100 mg/kg toabout 5 g/kg. DHA may be in the nutritional composition at about 500mg/kg to about 5 g/kg. DHA may be in the nutritional composition atabout 50 mg/kg to about 1 g/kg. For example, DHA may be in thenutritional composition at about 10 mg/kg, 20 mg/kg, 30 mg/kg, 40 mg/kg,50 mg/kg, 60 mg/kg, 70 mg/kg, 80 mg/kg, 90 mg/kg, 100 mg/kg, 110 mg/kg,120 mg/kg, 130 mg/kg, 140 mg/kg, 150 mg/kg, 160 mg/kg, 170 mg/kg, 180mg/kg, 190 mg/kg, 200 mg/kg, 210 mg/kg, 220 mg/kg, 230 mg/kg, 240 mg/kg,250 mg/kg, 260 mg/kg, 270 mg/kg, 280 mg/kg, 290 mg/kg, 300 mg/kg, 310mg/kg, 320 mg/kg, 330 mg/kg, 340 mg/kg, 350 mg/kg, 360 mg/kg, 370 mg/kg,380 mg/kg, 390 mg/kg, 400 mg/kg, 410 mg/kg, 420 mg/kg, 430 mg/kg, 440mg/kg, 450 mg/kg, 460 mg/kg, 470 mg/kg, 480 mg/kg, 490 mg/kg, 500 mg/kg,510 mg/kg, 520 mg/kg, 530 mg/kg, 540 mg/kg, 550 mg/kg, 560 mg/kg, 570mg/kg, 580 mg/kg, 590 mg/kg, 600 mg/kg, 610 mg/kg, 620 mg/kg, 630 mg/kg,640 mg/kg, 650 mg/kg, 660 mg/kg, 670 mg/kg, 680 mg/kg, 690 mg/kg, 700mg/kg, 710 mg/kg, 720 mg/kg, 730 mg/kg, 740 mg/kg, 750 mg/kg, 760 mg/kg,770 mg/kg, 780 mg/kg, 790 mg/kg, 800 mg/kg, 810 mg/kg, 820 mg/kg, 830mg/kg, 840 mg/kg, 850 mg/kg, 860 mg/kg, 870 mg/kg, 880 mg/kg, 890 mg/kg,900 mg/kg, 910 mg/kg, 920 mg/kg, 930 mg/kg, 940 mg/kg, 950 mg/kg, 960mg/kg, 970 mg/kg, 980 mg/kg, 990 mg/kg, 1 g/kg, 1.2 g/kg, 1.4 g/kg, 1.6g/kg, 1.8 g/kg, 2 g/kg, 2.2 g/kg, 2.4 g/kg, 2.6 g/kg, 2.8 g/kg, 3 g/kg,3.2 g/kg, 3.4 g/kg, 3.6 g/kg, 3.8 g/kg, 4 g/kg, 4.2 g/kg, 4.4 g/kg, 4.6g/kg, 4.8 g/kg, 5 g/kg, 5.2 g/kg, 5.4 g/kg, 5.6 g/kg, 5.8 g/kg, 6 g/kg,6.2 g/kg, 6.4 g/kg, 6.6 g/kg, 6.8 g/kg, 7 g/kg, 7.2 g/kg, 7.4 g/kg, 7.6g/kg, 7.8 g/kg, 8 g/kg, 8.2 g/kg, 8.4 g/kg, 8.6 g/kg, 8.8 g/kg, 9 g/kg,9.2 g/kg, 9.4 g/kg, 9.6 g/kg, 9.8 g/kg, or 10 g/kg.

(5) Combination of DHA and Lutein

In one embodiment, it has been unexpectedly found that when DHA andlutein are present in a composition according to the present disclosure,the bioavailability of DHA and lutein are improved upon ingestion overnutrient composition without the combination of DHA and lutein. Theimproved bioavailability allows DHA and lutein to be more readilyabsorbed into the body of the subject. Because the bioavailability ofDHA and lutein may be increased using the presently described processes,the level of DHA and lutein fortification required in the composition toprovide the desired nutritional benefit may be reduced. DHA and luteinmay be incorporated into the nutritional composition as recited in thevalues above.

c. Fat

The nutritional composition may comprise at least one fat. The fat maybe incorporated into the assembly. The fat may be present within thepremix, activated premix, and/or the protected premix. The fat may be amonoglyceride, a diglyceride or combination thereof (all hereafter“MDG”). Fat allows unique interactions with lipophilic compounds andproteins to form a water soluble, stable assembly. Accordingly, thepresence of the fat aids in assembly formation and subsequent stabilityof lipophilic containing assemblies within the composition, relative tocontrols without MDG and hydrophobic protein. The fat also enhances thebioavailability of the lipophilic compound, relative to controls withoutMDG or hydrophobic protein. Fat may also increase protection of thenutritional composition from dissolved oxygen. Non-limiting examples ofsuitable fats or sources thereof, in addition to the mixture ofmonoglycerides and diglycerides discussed above, for use in thenutritional composition described herein may be selected from the group,but is not limited to, coconut oil, fractionated coconut oil, soy oil,corn oil, olive oil, safflower oil, high oleic coconut oil, fractionatedcoconut oil, soy oil, corn oil, olive oil, safflower oil, high oleicsafflower oil, sunflower oil, high oleic sunflower oil, palm oil, palmkernel oil, palm olein, canola oil, marine oils, cottonseed oil, flaxseed oil, hemp seed oil, peanut oil, borage oil, algal oils, fungaloils, MCT oil (medium chain triglycerides) and combinations thereof.

(1) Monoglycerides and Diglycerides (MDG)

The fat of the nutritional composition may by MDG. MDG is thecombination of monoglycerides and diglycerides. MDG may be incorporatedinto the assembly. Incorporation of MDG as part of the assembly enhancesthe solubility of lipophilic compounds, thus enhancing itsbioavailability for treatment of health conditions, relative to controlswithout MDG or hydrophobic protein. It is hypothesized, without beingbound to a particular theory, that upon consumption, the MDG in theactivated premix or the MDG-protected premix, associate with thelipophilic compound and the hydrophobic protein making it easier for itto be incorporated into the micelles which are formed in the gut. Thus,the lipophilic compound may be more efficiently delivered to the brushboarder of the intestine and absorbed into the body. Additionally, it ishypothesized, without being bound to a particular theory, that MDG mayinteract with the hydrophobic protein and increase access of hydrophobicdomains within said protein. Accordingly, the lipophilic compound mayform more stable interactions with the newly accessible hydrophobicdomains of said protein.

MDG is a component with limited water solubility. When MDG is associatedwith the at least one hydrophobic protein and the at least onelipophilic compound, as in the assembly, its water solubility increasesignificantly. It is hypothesized, without being bound to any particulartheory, that the MDG that remains in the aqueous phase followingcentrifugation are likely associated with the assembly due to itslimited aqueous solubility. The increased aqueous solubility mayinvestigated by centrifugation, as listed above, wherein the aqueousphase following centrifugation exhibits a greater presence of MDGcompared to controls without MDG, lipophilic compound and at least onehydrophobic protein.

Diglycerides, which are commonly referred to as diacylglycerols (DAGs),are glycerides consisting of two fatty acid chains covalently bonded toa glycerol molecule through ester linkages. Monoglycerides are normalmetabolites in the body formed during the breakdown of triglycerides anddiglycerides. Non-limiting examples of commercial sources ofmonoglycerides and diglycerides include natural sources (e.g., animal(cow- or hog-derived) or vegetable, those which may be derived frompartially hydrogenated soy bean, sunflower, safflower and coconut oil)or synthetic sources. In some embodiments, the mixture of monoglyceridesand diglycerides may be derived from the hydrolysis of triglycerides insafflower oil. Monoglycerides may be in the MDG of the nutritionalcomposition at about 0.1 wt % to about 99.9 wt %, wherein the balance ofthe MDG is diglycerides. Monoglycerides may be in the MDG of thenutritional composition at about 10 wt % to about 80 wt %, wherein thebalance of the MDG is diglycerides. Monoglycerides may be in the MDG ofthe nutritional composition at about 25 wt % to about 50 wt %, whereinthe balance of the MDG is diglycerides. For example, monoglycerides maybe in the MDG of the nutritional composition at about 0.1 wt %, 0.5 wt%, 1 wt %, 1.5 wt %, 2 wt %, 2.5 wt %, 3 wt %, 3.5 wt %, 4 wt %, 4.5 wt%, 5 wt %, 5.5 wt %, 6 wt %, 6.5 wt %, 7 wt %, 7.5 wt %, 8 wt %, 8.5 wt%, 9 wt %, 9.5 wt %, 10 wt %, 10.5 wt %, 11 wt %, 11.5 wt %, 12 wt %,12.5 wt %, 13 wt %, 13.5 wt %, 14 wt %, 14.5 wt %, 15 wt %, 15.5 wt %,16 wt %, 16.5 wt %, 17 wt %, 17.5 wt %, 18 wt %, 18.5 wt %, 19 wt %,19.5 wt %, 20 wt %, 20.5 wt %, 21 wt %, 21.5 wt %, 22 wt %, 22.5 wt %,23 wt %, 23.5 wt %, 24 wt %, 24.5 wt %, 25 wt %, 25.5 wt %, 26 wt %,26.5 wt %, 27 wt %, 27.5 wt %, 28 wt %, 28.5 wt %, 29 wt %, 29.5 wt %,30 wt %, 30.5 wt %, 31 wt %, 31.5 wt %, 32 wt %, 32.5 wt %, 33 wt %,33.5 wt %, 34 wt %, 34.5 wt %, 35 wt %, 35.5 wt %, 36 wt %, 36.5 wt %,37 wt %, 37.5 wt %, 38 wt %, 38.5 wt %, 39 wt %, 39.5 wt %, 40 wt %,40.5 wt %, 41 wt %, 41.5 wt %, 42 wt %, 42.5 wt %, 43 wt %, 43.5 wt %,44 wt %, 44.5 wt %, 45 wt %, 45.5 wt %, 46 wt %, 46.5 wt %, 47 wt %,47.5 wt %, 48 wt %, 48.5 wt %, 49 wt %, 49.5 wt %, 50 wt %, 50.5 wt %,51 wt %, 51.5 wt %, 52 wt %, 52.5 wt %, 53 wt %, 53.5 wt %, 54 wt %,54.5 wt %, 55 wt %, 55.5 wt %, 56 wt %, 56.5 wt %, 57 wt %, 57.5 wt %,58 wt %, 58.5 wt %, 59 wt %, 59.5 wt %, 60 wt %, 60.5 wt %, 61 wt %,61.5 wt %, 62 wt %, 62.5 wt %, 63 wt %, 63.5 wt %, 64 wt %, 64.5 wt %,65 wt %, 65.5 wt %, 66 wt %, 66.5 wt %, 67 wt %, 67.5 wt %, 68 wt %,68.5 wt %, 69 wt %, 69.5 wt %, 70 wt %, 70.5 wt %, 71 wt %, 71.5 wt %,72 wt %, 72.5 wt %, 73 wt %, 73.5 wt %, 74 wt %, 74.5 wt %, 75 wt %,75.5 wt %, 76 wt %, 76.5 wt %, 77 wt %, 77.5 wt %, 78 wt %, 78.5 wt %,79 wt %, 79.5 wt %, 80 wt %, 80.5 wt %, 81 wt %, 81.5 wt %, 82 wt %,82.5 wt %, 83 wt %, 83.5 wt %, 84 wt %, 84.5 wt %, 85 wt %, 85.5 wt %,86 wt %, 86.5 wt %, 87 wt %, 87.5 wt %, 88 wt %, 88.5 wt %, 89 wt %,89.5 wt %, 90 wt %, 90.5 wt %, 91 wt %, 91.5 wt %, 92 wt %, 92.5 wt %,93 wt %, 93.5 wt %, 94 wt %, 94.5 wt %, 95 wt %, 95.5 wt %, 96 wt %,96.5 wt %, 97 wt %, 97.5 wt %, 98 wt %, 98.5 wt %, 99 wt %, 99.5 wt %,or 99.9%, wherein the balance of the MDG is diglycerides.

Non-limiting examples of suitable mixtures of monoglycerides anddiglycerides for use in the composition include Capmul GMO-40 (a MDGderived from high oleic safflower oil, available from AbitecCorporation, Columbus, Ohio), and Kirnol CE1089 (a MDG derived from cornoil, available from BASF, Ludwigshafen, Germany), RADIAMULS MG F038 (aMDG derived from high oleic sunflower oil, available from Oleon,Ertvelde, Belgium).

The MDG may be in the nutritional composition at about 140 mg/kg toabout 900 mg/kg. The MDG may be in the nutritional composition at about145 mg/kg to about 890 mg/kg. The nutritional composition may compriseMDG at about 140 mg/kg to about 700 mg/kg. The MDG may be in thenutritional composition at about 145 mg/kg to about 600 mg/kg. The MDGmay be in the nutritional composition at about 140 mg/kg to about 400mg/kg. For example, the MDG may be in the nutritional composition atabout 140 mg/kg, 145 mg/kg, 150 mg/kg, 155 mg/kg, 160 mg/kg, 165 mg/kg,170 mg/kg, 175 mg/kg, 180 mg/kg, 185 mg/kg, 190 mg/kg, 195 mg/kg, 200mg/kg, 205 mg/kg, 210 mg/kg, 215 mg/kg, 220 mg/kg, 225 mg/kg, 230 mg/kg,235 mg/kg, 240 mg/kg, 245 mg/kg, 250 mg/kg, 255 mg/kg, 260 mg/kg, 265mg/kg, 270 mg/kg, 275 mg/kg, 280 mg/kg, 285 mg/kg, 290 mg/kg, 295 mg/kg,300 mg/kg, 305 mg/kg, 310 mg/kg, 315 mg/kg, 320 mg/kg, 325 mg/kg, 330mg/kg, 335 mg/kg, 340 mg/kg, 345 mg/kg, 350 mg/kg, 355 mg/kg, 360 mg/kg,365 mg/kg, 370 mg/kg, 375 mg/kg, 380 mg/kg, 385 mg/kg, 390 mg/kg, 395mg/kg, 400 mg/kg, 405 mg/kg, 410 mg/kg, 415 mg/kg, 420 mg/kg, 425 mg/kg,430 mg/kg, 435 mg/kg, 440 mg/kg, 445 mg/kg, 450 mg/kg, 455 mg/kg, 460mg/kg, 465 mg/kg, 470 mg/kg, 475 mg/kg, 480 mg/kg, 485 mg/kg, 490 mg/kg,495 mg/kg, 500 mg/kg, 505 mg/kg, 510 mg/kg, 515 mg/kg, 520 mg/kg, 525mg/kg, 530 mg/kg, 535 mg/kg, 540 mg/kg, 545 mg/kg, 550 mg/kg, 555 mg/kg,560 mg/kg, 565 mg/kg, 570 mg/kg, 575 mg/kg, 580 mg/kg, 585 mg/kg, 590mg/kg, 595 mg/kg, 600 mg/kg, 605 mg/kg, 610 mg/kg, 615 mg/kg, 620 mg/kg,625 mg/kg, 630 mg/kg, 635 mg/kg, 640 mg/kg, 645 mg/kg, 650 mg/kg, 655mg/kg, 660 mg/kg, 665 mg/kg, 670 mg/kg, 675 mg/kg, 680 mg/kg, 685 mg/kg,690 mg/kg, 695 mg/kg, 700 mg/kg, 705 mg/kg, 710 mg/kg, 715 mg/kg, 720mg/kg, 725 mg/kg, 730 mg/kg, 735 mg/kg, 740 mg/kg, 745 mg/kg, 750 mg/kg,755 mg/kg, 760 mg/kg, 765 mg/kg, 770 mg/kg, 775 mg/kg, 780 mg/kg, 785mg/kg, 790 mg/kg, 795 mg/kg, 800 mg/kg, 805 mg/kg, 810 mg/kg, 815 mg/kg,820 mg/kg, 825 mg/kg, 830 mg/kg, 835 mg/kg, 840 mg/kg, 845 mg/kg, 850mg/kg, 855 mg/kg, 860 mg/kg, 865 mg/kg, 870 mg/kg, 875 mg/kg, 880 mg/kg,885 mg/kg, 890 mg/kg, 895 mg/kg, or 900 mg/kg.

In some embodiments, the nutritional composition may comprise about 0.1mg/kg to about 20 mg/kg of MDG and vitamin D. In some embodiments, thenutritional composition may comprise about 0.1 mg/kg to about 10 mg/kgof MDG and vitamin D.

(2) Palmitic Acid

The fat of the nutritional composition of the present invention may bepalmitic acid. Palmitic acid may be incorporated into the assembly.Incorporation of palmitic acid as part of the assembly enhances itssolubility, thus enhancing its bioavailability for treatment of healthconditions, relative to controls without MDG or hydrophobic protein.Palmitic acid may be used within the nutritional composition to maintainand supplement healthy nutrition, especially within infants.

Palmitic acid is a saturated fatty acid with 16 carbons in thelipophilic chain. Palmitic acid is an important energy source forgrowing infants. It is also a precursor to longer fatty acids that aresynthesized in the body. These longer fatty acids, such asdocosahexaenoic acid, are vital structural components in the brain,eyes, and central nervous system. As a consequence, palmitic acid is animportant nutrient in the healthy development of infants and babies.

Palmitic acid may be provided by HPAV oil, which comprises greater thanabout 22% palmitic acid, as a percentage of the total fatty acids in theHPAV oil. Examples of HPAV oil include, but are not limited to, palm oiland palm olein.

Palmitic acid is found in palm oil and palm olein, comprising over 30%of the total fatty acids in each oil. Palmitic acid is also found incocoa butter, cottonseed oil, and the oil of some other seed plants.

Palm oil is found in the pulp of the oil palm fruit. Palmitic acidcomprises from about 43% to about 45% of the fatty acids found in palmoil. Palm oil also comprises from about 37% to about 40% oleic acid,making it a good source of monounsaturated fat. Palm oil also containsthe essential fatty acid linoleic acid, which comprises from about 5% toabout 11% of the fatty acids in palm oil. Palm oil should not beconfused with palm kernel oil, which comes from the kernel (seed or pit)of the oil palm fruit. Palm kernel oil has a very different fatty acidprofile, comprising less than about 10% palmitic acid and primarilycomprising saturated fatty acids. Therefore, palm kernel oil is notconsidered a HPAV oil for the purpose of this description.

During refinement, raw palm oil is fractionated by crystallizing andseparating the liquid fraction from the solid fraction under controlledtemperatures. The liquid fraction is called palm olein. Palm oleincomprises somewhat less palmitic acid (from about 33% to about 40%) butmore oleic acid (from about 42% to about 48%) than does palm oil.

Palmitic acid may be provided by HPAV oil, which comprises greater thanabout 22% palmitic acid, as a percentage of the total fatty acids in theHPAV oil. Examples of HPAV oil include, but are not limited to, palm oiland palm olein.

Suppliers of suitable HPAV oils, such as palm oil or palm olein,include: Archer Daniels Midland, Decatur, Ill., USA; Fuji Vegetable Oil,Inc., Savannah, Ga., USA; and California Oils Corporation, Richmond,Calif., USA.

An HPAV oil may be provided in the nutritional composition as anindividual ingredient or in any combination with other materials orsources. For example, a HPAV oil may be provided in a mixture with otheroils such as canola oil or corn oil.

In some embodiments, it may be desirable to blend a protected premixcomprising the HPAV oil, a surfactant, and the MDG before blending theMDG oil or the HPAV oil with other ingredients of the nutritionalcomposition. In these embodiments, the HPAV oil and a surfactant aremixed with the MDG oil in the presence of heat or at ambient temperatureand, in some embodiments, with agitation to allow the HPAV oil todissolve or disperse in the MDG oil.

In a protected premix that contains HPAV oil, the HPAV oil is present inan amount relative to the amount of the MDG oil that will provideeffective amounts of the HPAV oil and the MDG oil in the infant formula.For example, the HPAV oil may be present in an amount of 15 g, about 20g, about 30 g, about 50 g, about 75 g, about 100 g, about 125 g, about150 g, about 175 g, or about 200 g, of the HPAV per 1 g of the MDG oilin the premix. Other ratios of the HPAV oil to the MDG oil are withinthe scope of the present disclosure, so long as the HPAV oil iscompletely or substantially dissolved or dispersed in the resultingpremix. Based on the disclosure herein, one skilled in the art maycalculate suitable ratios that will allow effective amounts of the HPAVoil and the MDG oil to be incorporated into the nutritional composition.

In human breast milk, about 70% of the palmitic acid is found in themiddle, or SN-2, location of the triglyceride. The SN-2 position isrelatively protected, and the palmitic acid remains bonded to theglycerol backbone until late in the digestive process. Palmitic acid inthe SN-2 position is thus in a form that is easily absorbed by theintestinal tract of the infant. However, for vegetable oils such as palmoil or palm olein, only about 9% of the palmitic acid is found in theSN-2 position. The rest of the palmitic acid in these oils is found inthe SN-1 or SN-3 locations on the glycerol backbone. Fatty acids in theSN-1 or SN-3 position are less protected, and are often cleaved off theglycerol backbone early in the digestive process. These free fatty acidsthen react with calcium in the gastric fluids, forming insolublecalcium-fatty acid soap complexes. These insoluble soap complexes, whichbind up both fatty acids and calcium, cannot be absorbed in theintestinal tract, and are instead excreted from the body. Research hasshown that infants who are fed formulas containing palm oil or palmolein do not absorb as much fat or calcium as infants who are fed breastmilk.

Palmitic acid may be synthetically prepared from fats that more closelymimic the fats in breast milk. These fats are synthesized to havesignificantly more palmitic acid in the SN-2 position than is found innatural vegetable oils. The nutrient composition may have palmitic acidin the SN-2 position at about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% ofthe total palmitic acid content of the nutrient composition.

Palmitic acid may be in the nutritional composition at about 0.5 g/kg toabout 35 g/kg. Palmitic acid may be in the nutritional composition atabout 1 g/kg to about 30 g/kg. Palmitic acid may be in the nutritionalcomposition at about 1 g/kg to about 20 g/kg. Palmitic acid may be inthe nutritional composition at about 1 g/kg to about 10 g/kg. Forexample, palmitic acid may be in the nutritional composition at about0.5 g/kg, 0.75 g/kg, 1 g/kg, 1.25 g/kg, 1.5 g/kg, 1.75 g/kg, 2 g/kg,2.25 g/kg, 2.5 g/kg, 2.75 g/kg, 3 g/kg, 3.25 g/kg, 3.5 g/kg, 3.75 g/kg,4 g/kg, 4.25 g/kg, 4.5 g/kg, 4.75 g/kg, 5 g/kg, 5.25 g/kg, 5.5 g/kg,5.75 g/kg, 6 g/kg, 6.25 g/kg, 6.5 g/kg, 6.75 g/kg, 7 g/kg, 7.25 g/kg,7.5 g/kg, 7.75 g/kg, 8 g/kg, 8.25 g/kg, 8.5 g/kg, 8.75 g/kg, 9 g/kg,9.25 g/kg, 9.5 g/kg, 9.75 g/kg, 10 g/kg, 10.25 g/kg, 10.5 g/kg, 10.75g/kg, 11 g/kg, 11.25 g/kg, 11.5 g/kg, 11.75 g/kg, 12 g/kg, 12.25 g/kg,12.5 g/kg, 12.75 g/kg, 13 g/kg, 13.25 g/kg, 13.5 g/kg, 13.75 g/kg, 14g/kg, 14.25 g/kg, 14.5 g/kg, 14.75 g/kg, 15 g/kg, 15.25 g/kg, 15.5 g/kg,15.75 g/kg, 16 g/kg, 16.25 g/kg, 16.5 g/kg, 16.75 g/kg, 17 g/kg, 17.25g/kg, 17.5 g/kg, 17.75 g/kg, 18 g/kg, 18.25 g/kg, 18.5 g/kg, 18.75 g/kg,19 g/kg, 19.25 g/kg, 19.5 g/kg, 19.75 g/kg, 20 g/kg, 20.25 g/kg, 20.5g/kg, 20.75 g/kg, 21 g/kg, 21.25 g/kg, 21.5 g/kg, 21.75 g/kg, 22 g/kg,22.25 g/kg, 22.5 g/kg, 22.75 g/kg, 23 g/kg, 23.25 g/kg, 23.5 g/kg, 23.75g/kg, 24 g/kg, 24.25 g/kg, 24.5 g/kg, 24.75 g/kg, 25 g/kg, 25.25 g/kg,25.5 g/kg, 25.75 g/kg, 26 g/kg, 26.25 g/kg, 26.5 g/kg, 26.75 g/kg, 27g/kg, 27.25 g/kg, 27.5 g/kg, 27.75 g/kg, 28 g/kg, 28.25 g/kg, 28.5 g/kg,28.75 g/kg, 29 g/kg, 29.25 g/kg, 29.5 g/kg, 29.75 g/kg, 30 g/kg, 30.25g/kg, 30.5 g/kg, 30.75 g/kg, 31 g/kg, 31.25 g/kg, 31.5 g/kg, 31.75 g/kg,32 g/kg, 32.25 g/kg, 32.5 g/kg, 32.75 g/kg, 33 g/kg, 33.25 g/kg, 33.5g/kg, 33.75 g/kg, 34 g/kg, 34.25 g/kg, 34.5 g/kg, 34.75 g/kg, or 35g/kg.

d. Protein

The nutritional composition may comprise at least one protein. The oneprotein may be incorporated into the assembly. The protein may bepresent within the activated premix and/or the protected premix. Theprotein uniquely interacts with the lipophilic compound and fat in thenutritional composition to form the water soluble, stable assemblydiscussed above. Accordingly, the presence of the protein aids inassembly formation and subsequent stability of lipophilic containingassemblies within the composition. The protein also enhances thebioavailability of the lipophilic compound, relative to controls withoutfat (e.g., MDG) or hydrophobic protein.

The protein may be intact protein, hydrolyzed protein or combinationsthereof. The protein may be a hydrophobic protein. Non-limiting examplesof suitable protein or sources thereof for use in the nutritionalcomposition include partially hydrolyzed (degree of hydrolysis less than25%) or non-hydrolyzed proteins (e.g., intact) or protein sources, whichmay be derived from any known or otherwise suitable source such as milk(e.g., casein, whey), animal (e.g., meat, fish), cereal (e.g., rice,corn) or combinations thereof. Non-limiting examples of such proteinsinclude milk protein isolates, milk protein concentrates as describedherein, such as whey protein concentrates, casein protein isolates, wheyprotein, caseinates, whole cow's milk, partially or completely defattedmilk, soy protein isolates, soy protein concentrates, and so forth.

In some embodiments, the nutritional composition includes a proteincomponent that consists of only intact and/or partially hydrolyzedprotein; that is, the protein component is free of any protein that hasa degree of hydrolysis of about 25% or more. In this context, the term“partially hydrolyzed protein” is used to refer to proteins having adegree of hydrolysis of less than about 25%, including less than about20%, including less than about 15%, including less than about 10%, andincluding proteins having a degree of hydrolysis of less than about 5%.The degree of hydrolysis is the extent to which peptide bonds are brokenby a hydrolysis method. The degree of protein hydrolysis for purposes ofcharacterizing the partially hydrolyzed protein component of theseembodiments is easily determined by one of ordinary skill in thecomposition arts by quantifying the amino nitrogen to total nitrogenratio (AN/TN) of the protein component of the selected composition. Theamino nitrogen component is quantified by USP titration methods fordetermining amino nitrogen content, while the total nitrogen componentis determined by the Tecator Kjeldahl method, all of which are wellknown methods to one of ordinary skill in the analytical chemistry art.

(1) Hydrophobic Protein

The protein of the nutritional composition may comprise at least onehydrophobic protein. The hydrophobic protein may be incorporated intothe assembly. Incorporation of the hydrophobic protein as part of theassembly enhances the solubility of lipophilic compounds, thus enhancingits bioavailability of the lipophilic compounds for treatment of healthconditions, relative to controls without MDG or hydrophobic protein. Itis hypothesized, without being bound to any particular theory, that thelipophilic compound's aqueous solubility is enhanced by associatingwithin the hydrophobic domains of said protein. The hydrophobic proteinmay be, but is not limited to, β-lactoglobulin, bovine lactoferrin,γ₂-casein, β-casein, α-lactalbumin, or combinations thereof. Thehydrophobic protein may be in the nutritional composition at about 1 g/Lto about 100 g/L. The hydrophobic protein may be in the nutritionalcomposition at about 10 g/L to about 80 g/L. The hydrophobic protein maybe in the nutritional composition at about 20 g/L to about 60 g/L. Thehydrophobic protein may be in the nutritional composition at about 30g/L to about 50 g/L. For example, the hydrophobic protein may be in thenutritional composition at about 1 g/L, 1.5 g/L, 2 g/L, 2.5 g/L, 3 g/L,3.5 g/L, 4 g/L, 4.5 g/L, 5 g/L, 5.5 g/L, 6 g/L, 6.5 g/L, 7 g/L, 7.5 g/L,8 g/L, 8.5 g/L, 9 g/L, 9.5 g/L, 10 g/L, 10.5 g/L, 11 g/L, 11.5 g/L, 12g/L, 12.5 g/L, 13 g/L, 13.5 g/L, 14 g/L, 14.5 g/L, 15 g/L, 15.5 g/L, 16g/L, 16.5 g/L, 17 g/L, 17.5 g/L, 18 g/L, 18.5 g/L, 19 g/L, 19.5 g/L, 20g/L, 20.5 g/L, 21 g/L, 21.5 g/L, 22 g/L, 22.5 g/L, 23 g/L, 23.5 g/L, 24g/L, 24.5 g/L, 25 g/L, 25.5 g/L, 26 g/L, 26.5 g/L, 27 g/L, 27.5 g/L, 28g/L, 28.5 g/L, 29 g/L, 29.5 g/L, 30 g/L, 30.5 g/L, 31 g/L, 31.5 g/L, 32g/L, 32.5 g/L, 33 g/L, 33.5 g/L, 34 g/L, 34.5 g/L, 35 g/L, 35.5 g/L, 36g/L, 36.5 g/L, 37 g/L, 37.5 g/L, 38 g/L, 38.5 g/L, 39 g/L, 39.5 g/L, 40g/L, 40.5 g/L, 41 g/L, 41.5 g/L, 42 g/L, 42.5 g/L, 43 g/L, 43.5 g/L, 44g/L, 44.5 g/L, 45 g/L, 45.5 g/L, 46 g/L, 46.5 g/L, 47 g/L, 47.5 g/L, 48g/L, 48.5 g/L, 49 g/L, 49.5 g/L, 50 g/L, 50.5 g/L, 51 g/L, 51.5 g/L, 52g/L, 52.5 g/L, 53 g/L, 53.5 g/L, 54 g/L, 54.5 g/L, 55 g/L, 55.5 g/L, 56g/L, 56.5 g/L, 57 g/L, 57.5 g/L, 58 g/L, 58.5 g/L, 59 g/L, 59.5 g/L, 60g/L, 60.5 g/L, 61 g/L, 61.5 g/L, 62 g/L, 62.5 g/L, 63 g/L, 63.5 g/L, 64g/L, 64.5 g/L, 65 g/L, 65.5 g/L, 66 g/L, 66.5 g/L, 67 g/L, 67.5 g/L, 68g/L, 68.5 g/L, 69 g/L, 69.5 g/L, 70 g/L, 70.5 g/L, 71 g/L, 71.5 g/L, 72g/L, 72.5 g/L, 73 g/L, 73.5 g/L, 74 g/L, 74.5 g/L, 75 g/L, 75.5 g/L, 76g/L, 76.5 g/L, 77 g/L, 77.5 g/L, 78 g/L, 78.5 g/L, 79 g/L, 79.5 g/L, 80g/L, 80.5 g/L, 81 g/L, 81.5 g/L, 82 g/L, 82.5 g/L, 83 g/L, 83.5 g/L, 84g/L, 84.5 g/L, 85 g/L, 85.5 g/L, 86 g/L, 86.5 g/L, 87 g/L, 87.5 g/L, 88g/L, 88.5 g/L, 89 g/L, 89.5 g/L, 90 g/L, 90.5 g/L, 91 g/L, 91.5 g/L, 92g/L, 92.5 g/L, 93 g/L, 93.5 g/L, 94 g/L, 94.5 g/L, 95 g/L, 95.5 g/L, 96g/L, 96.5 g/L, 97 g/L, 97.5 g/L, 98 g/L, 98.5 g/L, 99 g/L, 99.5 g/L, or100 g/L.

The hydrophobic protein may be characterized by its grand average ofhydropathicity index (GRAVY) value. Hydropathicity is used as defined inCreighton TE, Proteins: structures and molecular properties, 2ndedition, 1993, W.H. Freeman and Company, NY, N.Y., page 160. Thehydrophobic protein may have a GRAVY value of about −0.5 to about 0. Thehydrophobic protein may have a GRAVY value of about −0.4 to about 0. Thehydrophobic protein may have a GRAVY value of about −0.2 to about 0. TheGRAVY value of the hydrophobic protein is a measure of hydrophobicityand water solubility, and may be predictive of a protein's ability tobind a lipophilic molecule such as lutein. The GRAVY value may beobtained from protein data banks known within the art, as well asreferenced in Kyte et al., J. Mol. Biol. (1982) 157, 105-132,incorporated herein by reference in its entirety. The GRAVY value of thehydrophobic protein may be about −0.5, −0.498, −0.496, −0.494, −0.492,−0.49, −0.488, −0.486, −0.484, −0.482, −0.48, −0.478, −0.476, −0.474,−0.472, −0.47, −0.468, −0.466, −0.464, −0.462, −0.46, −0.458, −0.456,−0.454, −0.452, −0.45, −0.448, −0.446, −0.444, −0.442, −0.44, −0.438,−0.436, −0.434, −0.432, −0.43, −0.428, −0.426, −0.424, −0.422, −0.42,−0.418, −0.416, −0.414, −0.412, −0.41, −0.408, −0.406, −0.404, −0.402,−0.4, −0.398, −0.396, −0.394, −0.392, −0.39, −0.388, −0.386, −0.384,−0.382, −0.38, −0.378, −0.376, −0.374, −0.372, −0.37, −0.368, −0.366,−0.364, −0.362, −0.36, −0.358, −0.356, −0.354, −0.352, −0.35, −0.348,−0.346, −0.344, −0.342, −0.34, −0.338, −0.336, −0.334, −0.332, −0.33,−0.328, −0.326, −0.324, −0.322, −0.32, −0.318, −0.316, −0.314, −0.312,−0.31, −0.308, −0.306, −0.304, −0.302, −0.3, −0.298, −0.296, −0.294,−0.292, −0.29, −0.288, −0.286, −0.284, −0.282, −0.28, −0.278, −0.276,−0.274, −0.272, −0.27, −0.268, −0.266, −0.264, −0.262, −0.26, −0.258,−0.256, −0.254, −0.252, −0.25, −0.248, −0.246, −0.244, −0.242, −0.24,−0.238, −0.236, −0.234, −0.232, −0.23, −0.228, −0.226, −0.224, −0.222,−0.22, −0.218, −0.216, −0.214, −0.212, −0.21, −0.208, −0.206, −0.204,−0.202, −0.2, −0.198, −0.196, −0.194, −0.192, −0.19, −0.188, −0.186,−0.184, −0.182, −0.18, −0.178, −0.176, −0.174, −0.172, −0.17, −0.168,−0.166, −0.164, −0.162, −0.16, −0.158, −0.156, −0.154, −0.152, −0.15,−0.148, −0.146, −0.144, −0.142, −0.14, −0.138, −0.136, −0.134, −0.132,−0.13, −0.128, −0.126, −0.124, −0.122, −0.12, −0.118, −0.116, −0.114,−0.112, −0.11, −0.108, −0.106, −0.104, −0.102, −0.1, −0.098, −0.096,−0.094, −0.092, −0.09, −0.088, −0.086, −0.084, −0.082, −0.08, −0.078,−0.076, −0.074, −0.072, −0.07, −0.068, −0.066, −0.064, −0.062, −0.06,−0.058, −0.056, −0.054, −0.052, −0.05, −0.048, −0.046, −0.044, −0.042,−0.04, −0.038, −0.036, −0.034, −0.032, −0.03, −0.028, −0.026, −0.024,−0.022, −0.02, −0.018, −0.016, −0.014, −0.012, −0.01, −0.008, −0.006,−0.004, −0.002, or 0.

Additionally, it has been surprisingly found that hydrophobic proteinassociated with the assembly leads to a corresponding increase inlipophilic compound within the assembly, wherein hydrophobic proteincomprises from about 10 wt % to about 95 wt % of the assembly. Forexample, the hydrophobic protein may be in the assembly at about 10 wt%, 10.5 wt %, 11 wt %, 11.5 wt %, 12 wt %, 12.5 wt %, wt %, 13.5 wt %,14 wt %, 14.5 wt %, 15 wt %, 15.5 wt %, 16 wt %, 16.5 wt %, 17 wt %,17.5 wt %, 18 wt %, 18.5 wt %, 19 wt %, 19.5 wt %, 20 wt %, 20.5 wt %,21 wt %, 21.5 wt %, 22 wt %, 22.5 wt %, 23 wt %, 23.5 wt %, 24 wt %,24.5 wt %, 25 wt %, 25.5 wt %, 26 wt %, 26.5 wt %, 27 wt %, 27.5 wt %,28 wt %, 28.5 wt %, 29 wt %, 29.5 wt %, 30 wt %, 30.5 wt %, 31 wt %,31.5 wt %, 32 wt %, 32.5 wt %, wt %, 33.5 wt %, 34 wt %, 34.5 wt %, wt%, 35.5 wt %, wt %, 36.5 wt %, wt %, 37.5 wt %, 38 wt %, 38.5 wt %, 39wt %, 39.5 wt %, 40 wt %, 40.5 wt %, 41 wt %, 41.5 wt %, 42 wt %, 42.5wt %, 43 wt %, 43.5 wt %, 44 wt %, 44.5 wt %, 45 wt %, 45.5 wt %, 46 wt%, 46.5 wt %, 47 wt %, 47.5 wt %, 48 wt %, 48.5 wt %, 49 wt %, 49.5 wt%, 50 wt %, 50.5 wt %, 51 wt %, 51.5 wt %, 52 wt %, 52.5 wt %, wt %,53.5 wt %, 54 wt %, 54.5 wt %, 55 wt %, 55.5 wt %, 56 wt %, 56.5 wt %,57 wt %, 57.5 wt %, 58 wt %, 58.5 wt %, 59 wt %, 59.5 wt %, 60 wt %,60.5 wt %, 61 wt %, 61.5 wt %, 62 wt %, 62.5 wt %, 63 wt %, 63.5 wt %,64 wt %, 64.5 wt %, 65 wt %, 65.5 wt %, 66 wt %, 66.5 wt %, 67 wt %,67.5 wt %, 68 wt %, 68.5 wt %, 69 wt %, 69.5 wt %, 70 wt %, 70.5 wt %,71 AArt %, 71.5 wt %, 72 wt %, 72.5 wt %, 73 wt %, 73.5 wt %, 74 wt %,74.5 wt %, 75 wt %, 75.5 wt %, 76 wt %, 76.5 wt %, 77 wt %, 77.5 wt %,78 wt %, 78.5 wt %, 79 wt %, 79.5 wt %, 80 wt %, 80.5 wt %, 81 wt %,81.5 wt %, 82 wt %, 82.5 wt %, 83 wt %, 83.5 wt %, 84 wt %, 84.5 wt %,85 wt %, 85.5 wt %, 86 wt %, 86.5 wt %, 87 wt %, 87.5 wt %, 88 wt %,88.5 wt %, 89 wt %, 89.5 wt %, 90 wt %, 90.5 wt %, 91 wt %, 91.5 wt %,92 wt %, 92.5 wt %, wt %, 93.5 wt %, 94 wt %, 94.5 wt %, or 95 wt %.Additionally, the higher molecular weight of hydrophobic proteinassociated with the assembly correlates with an increase in lipophiliccompound within the assembly, wherein the minimum molecular weight ofprotein associated with the assembly may be about 5 kD to about 15 kD.For example, the minimum molecular weight of protein associated with theassembly may be about 5 kD, 5.2 kD, 5.4 kD, 5.6 kD, 5.8 kD, 6 kD, 6.2kD, 6.4 kD, 6.6 kD, 6.8 kD, 7 kD, 7.2 kD, 7.4 kD, 7.6 kD, 7.8 kD, 8 kD,8.2 kD, 8.4 kD, 8.6 kD, 8.8 kD, 9 kD, 9.2 kD, 9.4 kD, 9.6 kD, 9.8 kD, 10kD, 10.2 kD, 10.4 kD, 10.6 kD, 10.8 kD, 11 kD, 11.2 kD, 11.4 kD, 11.6kD, 11.8 kD, 12 kD, 12.2 kD, 12.4 kD, 12.6 kD, 12.8 kD, 13 kD, 13.2 kD,13.4 kD, 13.6 kD, 13.8 kD, 14 kD, 14.2 kD, 14.4 kD, 14.6 kD, 14.8 kD, or15 kD. In one embodiment, the minimum weight of protein associated withthe assembly is about 11.8 kD.

(a) β-casein

The hydrophobic protein of the nutritional composition may comprise atleast β-casein. β-casein may be incorporated into the assembly.Incorporation of β-casein as part of the assembly enhances thesolubility of lipophilic compounds, thus enhancing its bioavailabilityof the lipophilic compounds for treatment of health conditions, relativeto controls without MDG or hydrophobic protein. It has unexpectedly beenshown that β-casein is implicated in the delivery of certain lipophilicnutrients for absorption, digestion, or both. A2 β-casein is arelatively more hydrophobic form of β-casein (compared to A1 β-casein).The nutritional composition that replaces a portion of the inherentβ-casein with genetic variant A2 demonstrate enhanced bioavailability ofcertain lipophilic nutrients.

The majority of milk protein utilized in milk protein-containingproducts in the United States is from the cattle breed Holstein. Theterm “Holstein” as used herein should be understood to encompass theHolstein breed, the Friesian breed and cross-breeds of the two,so-called Holstein-Friesian cattle. Milk from Holstein cattle includesgenetic variant A1 as the predominant genetic variant. In contrast, milkfrom the Bos indicus breed has genetic variant A2 as the predominantgenetic variant. Similarly, milk from the Guernsey breed of Bos taurushas been shown to express high levels of β-casein variant A2 and lowlevels of other β-casein variants.

In certain embodiments, the nutritional composition may comprise proteinincluding 10 to 100% bovine β-casein. The bovine β-casein comprisesabout 50 to about 100% genetic variant A2. Thus, as a non-limitingexample, in a nutritional composition containing 10 grams of protein perserving, about 1-10 grams of that protein would be bovine-β-casein, andabout 0.5-10 grams of the protein would be genetic variant A2. Incertain exemplary embodiments the bovine β-casein comprises about 60% toabout 100%, including about 70% to about 100%, including about 80% toabout 100%, and including about 90% to about 100% by weight bovineβ-casein.

The bovine β-casein utilized in certain exemplary embodiments may befrom a single source, or may alternatively be provided by a combinationof sources. The bovine β-casein according to certain exemplaryembodiments will generally be found in milk protein isolates and milkprotein concentrates, but may also be found in other milk proteinsources such as whole milk, nonfat dry milk, milk protein concentrate,total milk protein, milk protein isolate, acid casein, calciumcaseinate, sodium caseinate, magnesium caseinate, purified β-casein, andcombinations thereof. It may be possible to purify a milk proteinisolate (or another milk protein source) containing unacceptably highlevels of less-desirable genetic variants (e.g., those other than A2).Non-limiting examples of purification methods useful for reducingunacceptably high levels of β-casein genetic variants include: apreparative chromatographic process (e.g., affinity chromatography, ionexchange chromatography, reversed phase chromatography) or by aselective salt precipitation (e.g., ammonium sulfate). Alternatively, amilk protein source (such as milk protein isolate or milk proteinconcentrate) containing milk sourced from primarily non-Holstein cattle,and hence, reduced quantities of genetic variants other than A2 may alsobe utilized in the nutritional composition and methods disclosed herein.

The term “milk protein concentrate” is generally used to refer to a milkprotein containing product that has had a considerable amount of theinherent water from ordinary milk removed and also has had inherent fatfrom the ordinary milk removed. The term “milk protein isolate” isgenerally used to refer to a type of milk-protein containing productthat has not only had a considerable amount of the inherent water fromordinary milk removed and inherent fat but also a certain amount ofinherent lactose removed. In most instances, milk protein isolates maybe considered to be a type of further purified milk protein concentrate.Certain manufacturers may use the term milk protein concentrate to referto milk-based protein products even if they contain at least 85 weight %protein.

The protein in the nutritional composition according to certainembodiments, may be provided by a single source of protein or acombination of protein sources. As previously discussed, about 10 toabout 100% by weight of the protein present in the nutritionalcomposition comprises bovine β-casein. The remaining portion of theprotein (e.g., 0-90% by weight of the total protein present in thenutritional composition) may be selected from one or more other sources.As discussed in more detail below, these additional sources of proteinare not particularly limited and may include one or more of soy protein,whey protein or any other protein source, including but not limited tothose discussed herein. Furthermore, it should be understood that thesource for the remaining portion of the protein could also be selectedfrom milk protein isolate, milk protein concentrate, caseinates, ornon-fat dry milk that does not meet the previously discussedrequirements as long as these components are not present in amounts sohigh as to violate other limitations on the amounts of genetic variantsdiscussed previously.

β-casein may be in the nutritional composition at about 0.2 g/L to about20 g/L. β-casein may be in the nutritional composition at about 0.5 g/Lto about 15 g/L. β-casein may be in the nutritional composition at about1 g/L to about 15 g/L. β-casein may be in the nutritional composition atabout 5 g/L to about 15 g/L. For example, β-casein may be in thenutritional composition at about 0.2 g/L, 0.4 g/L, 0.6 g/L, 0.8 g/L, 1g/L, 1.2 g/L, 1.4 g/L, 1.6 g/L, 1.8 g/L, 2 g/L, 2.2 g/L, 2.4 g/L, 2.6g/L, 2.8 g/L, 3 g/L, 3.2 g/L, 3.4 g/L, 3.6 g/L, 3.8 g/L, 4 g/L, 4.2 g/L,4.4 g/L, 4.6 g/L, 4.8 g/L, 5 g/L, 5.2 g/L, 5.4 g/L, 5.6 g/L, 5.8 g/L, 6g/L, 6.2 g/L, 6.4 g/L, 6.6 g/L, 6.8 g/L, 7 g/L, 7.2 g/L, 7.4 g/L, 7.6g/L, 7.8 g/L, 8 g/L, 8.2 g/L, 8.4 g/L, 8.6 g/L, 8.8 g/L, 9 g/L, 9.2 g/L,9.4 g/L, 9.6 g/L, 9.8 g/L, 10 g/L, 10.2 g/L, 10.4 g/L, 10.6 g/L, 10.8g/L, 11 g/L, 11.2 g/L, 11.4 g/L, 11.6 g/L, 11.8 g/L, 12 g/L, 12.2 g/L,12.4 g/L, 12.6 g/L, 12.8 g/L, 13 g/L, 13.2 g/L, 13.4 g/L, 13.6 g/L, 13.8g/L, 14 g/L, 14.2 g/L, 14.4 g/L, 14.6 g/L, 14.8 g/L, 15 g/L, 15.2 g/L,15.4 g/L, 15.6 g/L, 15.8 g/L, 16 g/L, 16.2 g/L, 16.4 g/L, 16.6 g/L, 16.8g/L, 17 g/L, 17.2 g/L, 17.4 g/L, 17.6 g/L, 17.8 g/L, 18 g/L, 18.2 g/L,18.4 g/L, 18.6 g/L, 18.8 g/L, 19 g/L, 19.2 g/L, 19.4 g/L, 19.6 g/L, 19.8g/L, or 20 g/L.

TABLE 2 Ratio of β-casein to Lipophilic Nutrients Table 1 Ratio ofβ-casein to Concentration, μM lipophilic nutrient, molar, Nutrient(approximate) approx. β-casein (~2.40 g/L) 102 N/A vitamin A (2029 IU/L)2.12 50 vitamin D (406 IU/L) 0.026 4000 vitamin E (10.1 IU/L) 0.016 6400vitamin K (54 μg/L) 0.120 900 Lutein (50 μg/L) 0.088 1200 ARA (150 mg/L)500 0.20 DHA (65 mg/L) 200 0.50

e. Optional Ingredients

The nutritional composition may further comprise other optionalingredients that may modify the physical, chemical, hedonic orprocessing characteristics of the products or serve as pharmaceutical oradditional nutritional components when used in the targeted population.Many such optional ingredients are known or otherwise suitable for usein other nutritional or pharmaceutical products and may also be used inthe nutritional composition described herein, provided that suchoptional ingredients are safe and effective for oral administration andare compatible with the essential and other ingredients in the selectedproduct form.

Non-limiting examples of such optional ingredients includecarbohydrates, preservatives, antioxidants, emulsifying agents, buffers,pharmaceutical actives, additional nutrients as described herein,colorants, flavors, thickening agents and stabilizers, and so forth.

Non-limiting examples of suitable carbohydrates or sources thereof foruse in the nutritional composition described herein may be selected fromselected from the group of: maltodextrin, hydrolyzed or modified starchor cornstarch, glucose polymers, corn syrup, corn syrup solids,rice-derived carbohydrates, glucose, fructose, lactose, high fructosecorn syrup, honey, sugar alcohols (e.g., maltitol, erythritol,sorbitol), and combinations thereof.

The nutritional composition may further comprise vitamins or relatednutrients, non-limiting examples of which may be selected from the groupof: vitamin A, vitamin K, thiamine, riboflavin, pyridoxine, vitamin B12,carotenoids (in addition to the lutein discussed above), niacin, folicacid, pantothenic acid, biotin, vitamin C, choline, inositol, salts, andderivatives thereof, and combinations thereof.

The nutritional composition may further comprise minerals, non-limitingexamples of which may be selected from the group of: phosphorus,magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum,chromium, selenium, chloride, and combinations thereof.

In some embodiments, the nutritional composition may comprise a compoundselected from the group of: hydroxyl methyl butyrate; leucine; betaalanine; epigallocatechin gallate; human milk oligosaccharides;prebiotics; probiotics; and combinations thereof.

The nutritional composition may also include one or more masking agentsto reduce or otherwise obscure bitter flavors and after taste. Suitablemasking agents may be selected from the group of: natural and artificialsweeteners, sodium sources such as sodium chloride, and hydrocolloids,such as guar gum, xanthan gum, carrageenan, gellan gum, and combinationsthereof. The amount of masking agent in the composition may varydepending upon the particular masking agent selected, other ingredientsin the composition, and other composition or product target variables.Such amounts, however, most typically range from at least about 0.1%,including from about 0.15% to about 3.0%, and also including from about0.18% to about 2.5%, by weight of the composition.

The nutritional composition may also optionally include one or morestabilizers. Appropriate stabilizers to use in formulation for thenutritional products include, but are not limited to, gum arabic, gumghatti, gum karaya, gum tragacanth, agar, furcellaran, locust bean gum,pectin, low methoxyl pectin, gelatin, microcrystalline cellulose, CMC(sodium carboxymethylcellulose), methylcellulose hydroxypropyl methylcellulose, hydroxypropyl cellulose, DATEM (diacetyl tartaric acid estersof mono- and diglycerides), dextran, CITREM (citric acid esters of mono-and diglycerides), and mixtures thereof.

(1) Choline

The optional ingredients of the nutritional composition may comprisecholine. Choline may act as both a nutrient within the composition, aswell as an agent that enhances assembly stability and formation. Cholinemay also lead to an increase in lipophilic concentration within theassembly. It is hypothesized, without being bound to any particulartheory, that the positive charge on choline interacts with negative sidechains of the hydrophobic protein (e.g., aspartic acid residues), andalters the 3D configuration of said protein. This may allow for moreaccess to the hydrophobic domains of the protein by the lipophiliccompound. Additionally, the use of choline may be lutein specific, inthat choline selectively increases the amount of lutein within theassembly, relative to other lipophilic compounds.

Choline may be added at different stages of the composition. The stagein which choline is added to the composition is critical to theformation of stable, water soluble assemblies. Specifically, it is moreadvantageous to add choline to the protein slurry, relative directly tothe premix/activated premix of MDG and lipophilic compound.

Choline acts as a source of methyl groups for the biosynthesis of othermethylated products. It is the precursor of the neurotransmitteracetylcholine. It has been proved that the administration of choline isbeneficial in patients suffering from any disorder related to defectivecholinergic neurotransmission.

Choline is also a major component, along with lecithin, of phospholipidsand sphingomyelin. By virtue of its fundamental functions in membranestructure, a choline deficiency causes a whole range of phospholipidabnormalities which express themselves clinically as fatty liver, kidneylesions (haemorrhagic renal necrosis) and impairment of lipoproteinmetabolism. With a diet deficient in choline, cholesterol esters andfats accumulate in the liver.

Choline chloride and bitartrate are mentioned in the US Code of FederalRegulations as nutrition/dietary supplements which have been accordedGRAS status (Generally Recognised As Safe).

Choline may be in the nutritional composition at about 5 mg/kg to about1 g/kg. Choline may be in the nutritional composition at about 50 mg/kgto about 1 g/kg. Choline may be in the nutritional composition at about100 mg/kg to about 1 g/kg. Choline may be in the nutritional compositionat about 500 mg/kg to about 1 g/kg. For example, choline may be in thenutritional composition at about 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg,25 mg/kg, 30 mg/kg, 35 mg/kg, 40 mg/kg, 45 mg/kg, 50 mg/kg, 55 mg/kg, 60mg/kg, 65 mg/kg, 70 mg/kg, 75 mg/kg, 80 mg/kg, 85 mg/kg, 90 mg/kg, 95mg/kg, 100 mg/kg, 105 mg/kg, 110 mg/kg, 115 mg/kg, 120 mg/kg, 125 mg/kg,130 mg/kg, 135 mg/kg, 140 mg/kg, 145 mg/kg, 150 mg/kg, 155 mg/kg, 160mg/kg, 165 mg/kg, 170 mg/kg, 175 mg/kg, 180 mg/kg, 185 mg/kg, 190 mg/kg,195 mg/kg, 200 mg/kg, 205 mg/kg, 210 mg/kg, 215 mg/kg, 220 mg/kg, 225mg/kg, 230 mg/kg, 235 mg/kg, 240 mg/kg, 245 mg/kg, 250 mg/kg, 255 mg/kg,260 mg/kg, 265 mg/kg, 270 mg/kg, 275 mg/kg, 280 mg/kg, 285 mg/kg, 290mg/kg, 295 mg/kg, 300 mg/kg, 305 mg/kg, 310 mg/kg, 315 mg/kg, 320 mg/kg,325 mg/kg, 330 mg/kg, 335 mg/kg, 340 mg/kg, 345 mg/kg, 350 mg/kg, 355mg/kg, 360 mg/kg, 365 mg/kg, 370 mg/kg, 375 mg/kg, 380 mg/kg, 385 mg/kg,390 mg/kg, 395 mg/kg, 400 mg/kg, 405 mg/kg, 410 mg/kg, 415 mg/kg, 420mg/kg, 425 mg/kg, 430 mg/kg, 435 mg/kg, 440 mg/kg, 445 mg/kg, 450 mg/kg,455 mg/kg, 460 mg/kg, 465 mg/kg, 470 mg/kg, 475 mg/kg, 480 mg/kg, 485mg/kg, 490 mg/kg, 495 mg/kg, 500 mg/kg, 505 mg/kg, 510 mg/kg, 515 mg/kg,520 mg/kg, 525 mg/kg, 530 mg/kg, 535 mg/kg, 540 mg/kg, 545 mg/kg, 550mg/kg, 555 mg/kg, 560 mg/kg, 565 mg/kg, 570 mg/kg, 575 mg/kg, 580 mg/kg,585 mg/kg, 590 mg/kg, 595 mg/kg, 600 mg/kg, 605 mg/kg, 610 mg/kg, 615mg/kg, 620 mg/kg, 625 mg/kg, 630 mg/kg, 635 mg/kg, 640 mg/kg, 645 mg/kg,650 mg/kg, 655 mg/kg, 660 mg/kg, 665 mg/kg, 670 mg/kg, 675 mg/kg, 680mg/kg, 685 mg/kg, 690 mg/kg, 695 mg/kg, 700 mg/kg, 705 mg/kg, 710 mg/kg,715 mg/kg, 720 mg/kg, 725 mg/kg, 730 mg/kg, 735 mg/kg, 740 mg/kg, 745mg/kg, 750 mg/kg, 755 mg/kg, 760 mg/kg, 765 mg/kg, 770 mg/kg, 775 mg/kg,780 mg/kg, 785 mg/kg, 790 mg/kg, 795 mg/kg, 800 mg/kg, 805 mg/kg, 810mg/kg, 815 mg/kg, 820 mg/kg, 825 mg/kg, 830 mg/kg, 835 mg/kg, 840 mg/kg,845 mg/kg, 850 mg/kg, 855 mg/kg, 860 mg/kg, 865 mg/kg, 870 mg/kg, 875mg/kg, 880 mg/kg, 885 mg/kg, 890 mg/kg, 895 mg/kg, 900 mg/kg, 905 mg/kg,910 mg/kg, 915 mg/kg, 920 mg/kg, 925 mg/kg, 930 mg/kg, 935 mg/kg, 940mg/kg, 945 mg/kg, 950 mg/kg, 955 mg/kg, 960 mg/kg, 965 mg/kg, 970 mg/kg,975 mg/kg, 980 mg/kg, 985 mg/kg, 990 mg/kg, 995 mg/kg, or 1 g/kg.

(2) Lecithin

The optional ingredients of the nutritional composition may compriselecithin. Lecithin may act as both a nutrient source within thecomposition, as well as an agent that enhances assembly stability andformation. Lecithin may also lead to an increase in lipophilicconcentration within the assembly. Lecithin is predominantly a mixtureof glycerol phospholipids (e.g., phosphatidylcholine,phosphatidylethanolamine and phosphatidylinositol). Phosphatidylcholineis typically the major glycerol phospholipid component. Lecithin mayalso contain other compounds such as free fatty acids, monoglycerides,diglycerides, triglycerides, glycolipids, and other lipid/fatty acidcontaining compounds. Lecithins are sometimes classified as glycerolphospholipids or phosphatides. This class of compounds has amphiphilicproperties and thus, emulsifying functionality.

Lecithin is typically added to liquid food products (includingnutritional compositions), so that the liquid products remainhomogeneous and does not separate. Lecithin is approved by the UnitedStates Food and Drug Administration for human consumption with thestatus “Generally Recognized As Safe.” Non-limiting examples of lecithinsuitable for use herein include egg lecithin, wheat lecithin, cornlecithin, soy lecithin, modified lecithin, and combinations thereof.Lecithin may be provided in deoiled or liquid form, orphosphatidylcholine enriched form. Additionally, lecithin may be derivedfrom sources that include, but are not limited to organic soy, canolaoil, nonfat dry milk, or whey protein.

In one embodiment, the nutritional powder may comprise deoiled lecithin,wherein the deoiled lecithin comprises 1% water; 3% triglycerides; 24%phosphatidylcholine; 20% phosphatidylethanolamine; 14%phosphatidylinositol; 7% phosphatidic acid; 8% minor phospholipids; 8%complexed sugars; and 15% glycolipids.

In another embodiment, the nutritional powder may comprise liquidlecithin, wherein the liquid lecithin comprises 1% water; 37%triglycerides; 16% phosphatidylcholine; 13% phosphatidylethanolamine;10% phosphatidylinositol; 5% phosphatidic acid; 2% minor phospholipids;5% complexed sugars; and 11% glycolipids.

Lecithin suitable for use herein may be obtained from any known orotherwise suitable nutrition source. Non-limiting examples include soylecithin from ADM Specialty Food Ingredients, Decatur, Ill., USA; soylecithin from Solae, LLC, St. Louis, Mo., USA; and soy lecithin fromAmerican Lecithin Company, Oxford, Conn., USA.

Lecithin may be in the nutritional composition at about 10 mg/kg toabout 5 g/kg. Lecithin may be in the nutritional composition at about 50mg/kg to about 5 g/kg. Lecithin may be in the nutritional composition atabout 100 mg/kg to about 1 g/kg. Lecithin may be in the nutritionalcomposition at about 500 mg/kg to about 1 g/kg. For example, lecithinmay be in the nutritional composition at about 10 mg/kg, 15 mg/kg, 20mg/kg, 25 mg/kg, 30 mg/kg, 35 mg/kg, 40 mg/kg, 45 mg/kg, 50 mg/kg, 55mg/kg, 60 mg/kg, 65 mg/kg, 70 mg/kg, 75 mg/kg, 80 mg/kg, 85 mg/kg, 90mg/kg, 95 mg/kg, 100 mg/kg, 105 mg/kg, 110 mg/kg, 115 mg/kg, 120 mg/kg,125 mg/kg, 130 mg/kg, 135 mg/kg, 140 mg/kg, 145 mg/kg, 150 mg/kg, 155mg/kg, 160 mg/kg, 165 mg/kg, 170 mg/kg, 175 mg/kg, 180 mg/kg, 185 mg/kg,190 mg/kg, 195 mg/kg, 200 mg/kg, 205 mg/kg, 210 mg/kg, 215 mg/kg, 220mg/kg, 225 mg/kg, 230 mg/kg, 235 mg/kg, 240 mg/kg, 245 mg/kg, 250 mg/kg,255 mg/kg, 260 mg/kg, 265 mg/kg, 270 mg/kg, 275 mg/kg, 280 mg/kg, 285mg/kg, 290 mg/kg, 295 mg/kg, 300 mg/kg, 305 mg/kg, 310 mg/kg, 315 mg/kg,320 mg/kg, 325 mg/kg, 330 mg/kg, 335 mg/kg, 340 mg/kg, 345 mg/kg, 350mg/kg, 355 mg/kg, 360 mg/kg, 365 mg/kg, 370 mg/kg, 375 mg/kg, 380 mg/kg,385 mg/kg, 390 mg/kg, 395 mg/kg, 400 mg/kg, 405 mg/kg, 410 mg/kg, 415mg/kg, 420 mg/kg, 425 mg/kg, 430 mg/kg, 435 mg/kg, 440 mg/kg, 445 mg/kg,450 mg/kg, 455 mg/kg, 460 mg/kg, 465 mg/kg, 470 mg/kg, 475 mg/kg, 480mg/kg, 485 mg/kg, 490 mg/kg, 495 mg/kg, 500 mg/kg, 505 mg/kg, 510 mg/kg,515 mg/kg, 520 mg/kg, 525 mg/kg, 530 mg/kg, 535 mg/kg, 540 mg/kg, 545mg/kg, 550 mg/kg, 555 mg/kg, 560 mg/kg, 565 mg/kg, 570 mg/kg, 575 mg/kg,580 mg/kg, 585 mg/kg, 590 mg/kg, 595 mg/kg, 600 mg/kg, 605 mg/kg, 610mg/kg, 615 mg/kg, 620 mg/kg, 625 mg/kg, 630 mg/kg, 635 mg/kg, 640 mg/kg,645 mg/kg, 650 mg/kg, 655 mg/kg, 660 mg/kg, 665 mg/kg, 670 mg/kg, 675mg/kg, 680 mg/kg, 685 mg/kg, 690 mg/kg, 695 mg/kg, 700 mg/kg, 705 mg/kg,710 mg/kg, 715 mg/kg, 720 mg/kg, 725 mg/kg, 730 mg/kg, 735 mg/kg, 740mg/kg, 745 mg/kg, 750 mg/kg, 755 mg/kg, 760 mg/kg, 765 mg/kg, 770 mg/kg,775 mg/kg, 780 mg/kg, 785 mg/kg, 790 mg/kg, 795 mg/kg, 800 mg/kg, 805mg/kg, 810 mg/kg, 815 mg/kg, 820 mg/kg, 825 mg/kg, 830 mg/kg, 835 mg/kg,840 mg/kg, 845 mg/kg, 850 mg/kg, 855 mg/kg, 860 mg/kg, 865 mg/kg, 870mg/kg, 875 mg/kg, 880 mg/kg, 885 mg/kg, 890 mg/kg, 895 mg/kg, 900 mg/kg,905 mg/kg, 910 mg/kg, 915 mg/kg, 920 mg/kg, 925 mg/kg, 930 mg/kg, 935mg/kg, 940 mg/kg, 945 mg/kg, 950 mg/kg, 955 mg/kg, 960 mg/kg, 965 mg/kg,970 mg/kg, 975 mg/kg, 980 mg/kg, 985 mg/kg, 990 mg/kg, 995 mg/kg, 1g/kg, 1.2 g/kg, 1.4 g/kg, 1.6 g/kg, 1.8 g/kg, 2 g/kg, 2.2 g/kg, 2.4g/kg, 2.6 g/kg, 2.8 g/kg, 3 g/kg, 3.2 g/kg, 3.4 g/kg, 3.6 g/kg, 3.8g/kg, 4 g/kg, 4.2 g/kg, 4.4 g/kg, 4.6 g/kg, 4.8 g/kg, or 5 g/kg.

4. METHOD OF MANUFACTURING

To prepare the nutritional composition having improved bioavailabilityof a lipophilic compound contained therein, a method in accordance withthe disclosure may be utilized. The method may comprise the steps of:forming a premix; adding the premix to a solution to form an activatedpremix or a protected premix, and adding the activated premix to thecomposition, wherein the assembly may be at least formed in theactivated premix.

a. Premix

In some embodiments, a premix comprising MDG is provided. The premix maycomprise MGD alone, MGD and at least one lipophilic compound, as well asother optional ingredients, as described above. The premix may beactivated. The premix may be heated prior to being activated. In someembodiments, the premix is heated to about 85° F. for 30 min. In someembodiments, the premix is heated to 120° F. for about 10 minutes. Oneof skill would appreciate that the premix composition may affect thetemperature and time that is needed to heat said premix. The premix maybe protected, for example by surfactants.

(1) Activated Premix

The premix may be added to an aqueous solution, for example to aprotein-in-water slurry, to form an “activated premix.” The activatedpremix is then added to the composition which may comprise at least onelipophilic compound.

Notably, to form an “activated premix,” the premix may not be added to aquantity of oil that is sufficient to cause the MDG to disassociate fromthe assembly in the oil prior to the addition of the premix into theaqueous solution. In this instance, the activate premix may be protectedto limit dissociation of MDG and lipophilic compound from each other.

When an “activated premix” is subsequently added to a fat-containingsolution, at least a portion of the MDG is not disassociated from theassembly by the fat in the fat-containing solution. To determine theamount of the monoglycerides and diglycerides that are not disassociatedin the fat, the “Disassociated MDG (monoglyceride and diglyceride) Test”may be utilized.

The Disassociated MDG Test is as follows. The fat-containing solution issubjected to high speed centrifugation (31,000×g; 20° C.; 4 h), asdescribed above. The resulting aqueous fraction is subjected to HPLCanalysis for monoglycerides and diglycerides content as described in“Determination of food emulsifiers in commercial additives and foodproducts by liquid chromatography/atmospheric-pressure chemicalionization mass spectrophotometry,” by M. Suman et al., Journal ofChromatography A, 1216 (2009) 3758-3766. The level of monoglycerides anddiglycerides detected in the aqueous fraction is compared to the levelthat was added to the fat-containing solution to arrive at thepercentage of the mixture of monoglycerides and diglycerides that is notdisassociated by the fat.

Using the Disassociated MDG Test, one may determine the impact ofutilizing an activated premix versus a premix that has not beenactivated in a composition. This may be accomplished by comparing theamount of monoglycerides and diglycerides that is present in the aqueousfraction of a composition comprising an activated premix, with theamount of monoglycerides and diglycerides that is present in the aqueousfraction of the same composition in which the premix has not beenactivated. The aqueous fraction of the composition comprising theactivated premix will typically contain a higher amount ofmonoglycerides and diglycerides than the same composition in which thepremix has not been activated.

The difference between the content of monoglycerides and diglycerides inthe aqueous fraction of the composition comprising the activated premix,and the content of monoglycerides and diglycerides in the aqueousfraction of the same composition comprising a premix that has not beenactivated, may be quantified. In some embodiments, the content ofmonoglycerides and diglycerides in the aqueous fraction of thecomposition comprising the activated premix may be at least about 5%,10% or 15% higher than in the same composition that comprises a premixthat has not been activated.

(b) Variations of Manufacture

The premix, activated premix, and composition may comprise differentembodiments, as some are listed below. It should be noted that thenutritional composition is not limited to the representative embodimentslisted below.

In some embodiments, a premix comprising the lipophilic compound and MDGis provided. To make the premix, the lipophilic compound may be mixedwith the MDG in the presence of heat or at ambient temperature, and insome embodiments, with agitation to allow the lipophilic compound todissolve into the monoglycerides and diglycerides. The premix is addedto an aqueous solution, for example to a protein-in-water slurry, toform an activated premix, which contains the assembly. The activatedpremix is then added to the composition.

In some embodiments, a premix comprising the lipophilic compound,surfactant, MDG is provided. To make the premix, the lipophilic compoundmay be mixed with the MDG in the presence of heat or at ambienttemperature, and in some embodiments, with agitation to allow thelipophilic compound to dissolve into the mixture of monoglycerides anddiglycerides. The premix is added to a fat-containing solution, such asa protein-in-fat slurry, to form an “protected premix”, which containsthe assembly. The “protected premix” is then added to the composition.

Without wishing to be bound by theory, it is believed that thesurfactant combines with the lipophilic compound, monoglycerides anddiglycerides, to form a microemulsion when the premix is added to thefat-containing solution. Thus, in a “protected premix,” themonoglycerides and diglycerides may be protected from disassociating inthe fat-containing solution.

The activated premix or protected premix may be added to any otheringredients of the nutritional or pharmaceutical composition at anyuseful point during the manufacturing process. In some embodiments, thenutritional or pharmaceutical composition may then be dried to form apowdered composition using any methods known in the art. By way ofexample, nutritional powders may be prepared by preparing at least twoslurries that will later be blended together (and further may be blendedwith the activated premix), heat treated, standardized, heat treated asecond time, evaporated to remove water, and spray dried to form apowdered nutritional composition.

Slurries to which the activated premix or protected premix may be addedinclude a carbohydrate-mineral (CHO-MIN) slurry and a protein-in-oil(PIO) slurry. Initially, the CHO-MIN slurry may be formed by dissolvingselected carbohydrates (e.g., lactose, galactooligosaccharides, etc.) inheated water with agitation, followed by the addition of minerals (e.g.,potassium citrate, magnesium chloride, potassium chloride, sodiumchloride, choline chloride, etc.). The resulting CHO-MIN slurry may thenbe held with continued heat and moderate agitation until it is laterblended with the other prepared slurries.

The protein-in-oil (PIO) slurry may be formed by heating and mixing theoil (e.g., high oleic safflower oil, soybean oil, coconut oil,monoglycerides, etc.) and emulsifier (e.g., soy lecithin), and thenadding oil soluble vitamins, protein (e.g., milk protein concentrate,milk protein hydrolysate, etc.), carrageenan (if any), calcium carbonateor tricalcium phosphate (if any), and arachidonic acid (ARA) oil anddocosahexaenoic acid (DHA) oil (if any) with continued heat andagitation. The resulting PIO slurry may be held with continued heat andmoderate agitation until it is later blended with the other preparedslurries.

Water is heated and then combined with the CHO-MIN slurry, nonfat milk(if any), and the PIO slurry under adequate agitation. The pH of theresulting blend is adjusted to 6.6-7.0, and the blend is held undermoderate heated agitation. ARA oil and DHA oil is added at this stage insome embodiments. In some embodiments, the activated premix or protectedpremix is blended with one or more of the heated water, CHO-MIN slurry,nonfat milk (if any), and the PIO slurry. Alternatively, the activatedpremix or protected premix may be blended into the composition afterfurther processing and immediately prior to drying of the composition.

The composition may then be subjected to high-temperature short-time(HTST) processing, during which the composition is heat treated,emulsified and homogenized, and then cooled. Water soluble vitamins andascorbic acid are added, the pH is adjusted to the desired range ifnecessary, flavors (if any) are added, and water is added to achieve thedesired total solid level. In some embodiments, the emulsions may thenbe further diluted, and subsequently processed and packaged asready-to-drink or concentrated liquids. In some embodiments, theemulsions are evaporated, heat treated and subsequently processed andpackaged as reconstitutable powder, e.g., spray dried, dry blended,agglomerated.

The spray dried powdered nutritional composition or dry blended powderednutritional composition may be prepared by any collection of known orotherwise effective techniques, suitable for making and formulating anutritional powder. For example, the spray drying step may likewiseinclude any spray drying technique that is known for or otherwisesuitable for use in the production of nutritional powders. Manydifferent spray drying methods and techniques are known for use in thenutrition field, all of which are suitable for use in the manufacture ofthe spray dried powdered nutritional composition. Following drying, thefinished powder may be packaged into suitable containers.

5. METHODS OF USE

The nutritional composition may be packaged and sealed in single ormulti-use containers, and then stored under ambient conditions or underrefrigeration for up to 36 months or longer, more typically from about 6months to about 24 months. For multi-use containers, these packages maybe opened and then covered for repeated use by the ultimate user. Forliquid embodiments, the opened and subsequently recovered package istypically stored under refrigerated conditions, and the contents usedwithin about 7 days. For powdered embodiments, the opened andsubsequently recovered package may be typically stored under ambientconditions (e.g., avoid extreme temperatures) and the contents usedwithin about one month. Non-limiting examples of ways in which thepresent formulas may be utilized include use as the following products,use as: a beverage, e.g. a coffee beverage, a cocoa or chocolatebeverage, a malted beverage, a fruit or juice beverage, a carbonatedbeverage, a soft drink, or a milk based beverage; a performancenutrition product, e.g. a performance nutrition bar, powder orready-to-drink beverage; a medical nutrition product; a dairy product,e.g. a milk drink, a yogurt or other fermented dairy product; an icecream product; a confectionary product, e.g. a chocolate product; afunctional food or beverage, e.g. a slimming product, a fat burningproduct, a product for improving mental performance or preventing mentaldecline, or a skin improving product. A beverage according to theinvention may be in the form of a powder or liquid concentrate to bemixed with a suitable liquid, e.g. water or milk, before consumption; ora ready-to-drink beverage.

In embodiments directed toward enhancing the absorption of thelipophilic nutrient, the lipophilic nutrient is administered to thesubject in a nutritional composition comprising the lipophilic nutrientand a lipophilic MDG carrier oil of the nutritional composition. Theabsorption of the lipophilic nutrient may be enhanced in one or moremeasurable ways. The absorption of the lipophilic nutrient may beenhanced by increasing the maximum plasma concentration of the nutrientin the blood of the ingesting subject. The absorption of the lipophilicnutrient may be enhanced by prolonging the plasma absorption of thenutrient in the blood of the ingesting subject over a period of days orweeks. The absorption of the lipophilic nutrient may be enhanced byincreasing the total plasma concentration of the nutrient in the bloodof the ingesting subject over a period of days or weeks.

In embodiments in which the lipophilic compound is lutein, the powderednutritional composition including the lutein may be administered toimprove age-related macular degeneration and other retinal diseases anddisorders. Although in some embodiments, the methods of the presentdisclosure may be directed to individuals who have age-related maculardegeneration or other retinal diseases and disorders, the methods of thepresent disclosure as described herein are also intended in someembodiments to include the use of such methods in “at risk” individuals,including individuals unaffected by or not otherwise afflicted withage-related macular degeneration or other retinal diseases or disorders,for the purpose of preventing, minimizing, or delaying the developmentof such diseases and conditions over time. For such prevention purposes,the methods of the present disclosure preferably include continuous,daily administration of the composition as described herein. Suchpreventive methods may be directed at adults or others, particularlyolder adults, who are susceptible to developing age-related maculardegeneration or other retinal diseases and disorders due to hereditaryconsiderations, environmental considerations, and the like.

Composition including Vitamin D may be administered to subjects topromote healthy bone development, reduce Vitamin D deficiency, increasebone strength, preserve or increase bone mineralization, and preserve orincrease bone mineral density. The composition including Vitamin D mayalso be administered to subjects suffering from or at risk for sufferingfrom diseases and conditions associated with inadequate bonemineralization, such as rickets, osteomalacia, osteoporosis, osteopenia,and bone fractures. The subjects receiving the composition with VitaminD may include, but are not limited to, pregnant and post-natal women,infants, children, adolescents, adults, post-menopausal women, and olderadults. In some embodiments, the methods of the present disclosure aredirected to pregnant or post-natal women, to promote healthy bonedevelopment in the developing fetus or infant. In some embodiments, themethods of the present disclosure are directed to infants and children,to promote healthy bone development and prevent rickets or bonefractures in the growing child. In some embodiments, the methods of thepresent disclosure are directed to adolescents and adults, to promoteincreased bone mineralization and bone mineral density. In someembodiments, the methods of the present disclosure are directed topost-menopausal women, to prevent or slow the onset of osteoporosis. Insome embodiments, the methods of the present disclosure are directed toadults who have osteomalacia, osteoporosis, osteopenia, or other bonediseases and disorders. In some embodiments, the methods of the presentdisclosure are directed to “at risk” individuals, including individualsunaffected by or not otherwise afflicted by osteomalacia, osteoporosis,osteopenia, loss of balance and falling, bone fractures, or other bonediseases or disorders, for the purpose of preventing, minimizing, ordelaying the development of such diseases and conditions over time. Forsuch development, maintenance, and prevention purposes, the methods ofthe present disclosure preferably include continuous, dailyadministration of the composition as described herein. Such development,maintenance, and preventive methods are directed at subjects such aspregnant and postnatal women, infants, children, adolescents, adults,post-menopausal women, and older adults. Preventative methods areparticularly directed to older adults who are susceptible to developingage-related osteoporosis, osteopenia, loss of balance or falling, brokenbones, or other diseases and disorders associated with inadequateVitamin D consumption or inadequate bone mineralization.

In these embodiments, a subject desirably consumes servings of thecomposition that provide an effective amount of Vitamin D per day.Effective amounts of Vitamin D range from about 50 IU to about 7,500 IUper day, including from about 100 IU per day to about 5,000 IU per day,including from about 200 IU per day to about 2,500 IU per day, includingfrom about 250 IU per day to about 1,500 IU per day, including fromabout 400 IU per day to about 1,000 IU per day, and including from about500 IU per day to about 800 IU per day.

In embodiments in which the lipophilic nutrient is Vitamin E, thenutritional composition including Vitamin E may be administered tosubjects to provide antioxidants, promote cardiovascular health, andprevent or reduce the risks of some types of cancer. The subjectsreceiving the nutritional composition with Vitamin E may include, butare not limited to, pregnant and post-natal women, infants, children,and adults. In some embodiments, the methods of the present disclosuremay be directed to pregnant or post-natal women to promote healthydevelopment in the developing fetus or infant. In some embodiments, themethods of the present disclosure may be directed to infants andchildren to promote healthy development in the growing child. In someembodiments, the methods of the present disclosure may be directed toadults who have cardiovascular disease. In some embodiments, the methodsof the present disclosure are also directed to “at risk” individuals,including individuals unaffected by or not otherwise afflicted withcancer, cardiovascular disease, or other diseases or disorders, for thepurpose of preventing, minimizing, or delaying the development of suchdiseases and conditions over time. For such development, maintenance,and prevention purposes, the methods of the present disclosurepreferably include continuous, daily administration of the nutritionalcomposition as described herein. Such development, maintenance, andpreventive methods may be directed at subjects such as pregnant andpostnatal women, infants, children, and adults, particularly olderadults who are susceptible to developing cancer, cardiovascular disease,or other diseases and disorders due to hereditary considerations,environmental considerations, and the like.

In embodiments in which the lipophilic nutrient is DHA, the nutritionalcomposition including DHA may be administered to subjects to promotehealthy eye, brain, and central nervous system development, maintainoverall brain health, prevent or reduce age-related decline in mentalfunctioning, and reduce cardiovascular and other diseases and disorders.The subjects receiving the nutritional composition with DHA may include,but are not limited to, pregnant and post-natal women, infants,children, and adults. In some embodiments, the methods of the presentdisclosure may be directed to pregnant or post-natal women to promoteeye, brain, and central nervous system development in the developingfetus or infant. In some embodiments, the methods of the presentdisclosure may be directed to infants and children to promote eye,brain, and central nervous system development in the growing child. Insome embodiments, the methods of the present disclosure may be directedto adults to maintain overall brain health, prevent or reduceage-related decline in mental functioning, and reduce cardiovascular andother diseases and disorders. In some embodiments, the methods of thepresent disclosure are also directed to “at risk” individuals, includingindividuals unaffected by or not otherwise afflicted with age-relateddeclines in mental functioning or cardiovascular disease, for thepurpose of preventing, minimizing, or delaying the development of suchdiseases and conditions over time. For such development, maintenance,and prevention purposes, the methods of the present disclosurepreferably include continuous, daily administration of the nutritionalcomposition as described herein. Such development, maintenance, andpreventive methods may be directed at subjects such as pregnant andpostnatal women, infants, children, and adults, particularly olderadults who are susceptible to age-related declines in mentalfunctioning, cardiovascular disease, or other diseases and disorders dueto hereditary considerations, environmental considerations, and thelike.

The nutritional composition including HPAV oil and an activated MDG oilcomponent or a protected premix may be administered to infants toenhance palmitic acid absorbance, promote healthy bone development, andmaintain healthy bone mineral density. The subjects receiving thecomposition with an HPAV oil and an activated MDG oil component or aprotected premix include, but are not limited to, pre-term and terminfants. In some embodiments, the methods of the present disclosure maybe directed to infants, to promote healthy bone development and maintainhealthy bone mineral density in the growing infant. For such developmentand maintenance purposes, the methods of the present disclosurepreferably include continuous, daily administration of the nutritionalcomposition as described herein. Such development and maintenancemethods may be directed at subjects such as preterm and term infants.

In these embodiments, an infant desirably consumes servings of thenutritional composition that provide effective amounts of the HPAV oilper day. Effective amounts of the HPAV oil range from about 0.6 g toabout 18 g per day, including from about 1 g per day to about 15 g perday, including from about 2.5 g per day to about 12.5 g per day,including from about 5 g per day to about 10 g per day, and includingfrom about 6 g per day to about 8 g per day.

The nutritional composition including DHA and lutein may be administeredto subjects to promote healthy eye and retina development, maintainoverall eye health, and prevent or reduce age-related maculardegeneration and other diseases and disorders of the eye. The subjectsreceiving the nutritional composition with DHA and lutein may include,but are not limited to, pregnant and post-natal women, infants,children, and adults. In some embodiments, the methods of the presentdisclosure may be directed to pregnant or post-natal women, to promoteeye and retina development and healthy vision in the developing fetus orinfant. In some embodiments, the methods of the present disclosure maybe directed to infants, to promote eye and retina development and visualacuity in the growing child. In some embodiments, the methods of thepresent disclosure may be directed to children and adults to improvevisual acuity, improve photostress recovery, or reduce sensitivity toglare. In some embodiments, the methods of the present disclosure may bedirected to adults who have age-related macular degeneration, glaucoma,cataracts, lacrimal keratoconjunctivitis, or other diseases anddisorders of the eye. In some embodiments, the methods of the presentdisclosure are also directed to “at risk” individuals, includingindividuals unaffected by or not otherwise afflicted with age-relatedmacular degeneration, glaucoma, cataracts, lacrimalkeratoconjunctivitis, diabetic retinopathy, or other diseases ordisorders of the eye, for the purpose of preventing, minimizing, ordelaying the development of such diseases and conditions over time. Forsuch development, maintenance, and prevention purposes, the methods ofthe present disclosure preferably include continuous, dailyadministration of the nutritional composition as described herein. Suchdevelopment, maintenance, and preventive methods may be directed atsubjects such as pregnant and postnatal women, infants, children, andadults, particularly older adults who are susceptible to developingage-related macular degeneration or other diseases and disorders of theeye due to hereditary considerations, environmental considerations, andthe like.

In these embodiments, a subject desirably consumes servings of thecomposition that provide effective amounts of DHA and lutein per day.Effective amounts of DHA range from about 5 mg to about 10 g per day,including from about 10 mg per day to about 1 g per day, including fromabout 20 mg per day to about 500 mg per day, including from about 40 mgper day to about 200 mg per day, and including from about 80 mg per dayto about 150 mg per day. Effective amounts of lutein range from about 5μg to about 10 mg per day, including from about 10 μg per day to about 5mg per day, including from about 25 μg per day to about 1 mg per day,including from about 50 mg per day to about 500 μg per day, andincluding from about 100 mg per day to about 250 mg per day.

In these embodiments, an individual desirably consumes at least oneserving of the nutritional composition daily, and in some embodiments,may consume two, three, or even more servings per day. Each serving isdesirably administered as a single undivided dose, although the servingmay also be divided into two or more partial or divided servings to betaken at two or more times during the day. The methods of the presentdisclosure include continuous day after day administration, as well asperiodic or limited administration, although continuous day after dayadministration is generally desirable. The nutritional composition maybe for use in infants, children, and adults.

The nutritional composition has multiple aspects, illustrated by thefollowing non-limiting examples.

5. EXAMPLES

It is understood that the foregoing detailed description andaccompanying examples are merely illustrative and are not to be taken aslimitations upon the scope of the invention, which is defined solely bythe appended claims and their equivalents.

Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art. Such changes and modifications,including without limitation those relating to the chemical structures,substituents, derivatives, intermediates, syntheses, compositions, ormethods of use of the invention, may be made without departing from thespirit and scope thereof.

The following examples illustrate specific embodiments and/or featuresof the nutritional composition comprising an activated premix and thecomposition comprising a protected premix. The examples are given solelyfor the purpose of illustration and are not to be construed aslimitations of the present disclosure, as many variations thereof arepossible without departing from the spirit and scope of the disclosure.All exemplified amounts are weight percentages based upon the totalweight of the composition, unless otherwise specified.

The exemplified composition may be prepared in accordance with themanufacturing methods described herein, such that each exemplifiednutritional or pharmaceutical composition has improved lipophiliccompound bioavailability.

The following details may be applied to the below-listed Examples whenappropriate. Changes outside of the details listed below are recitedwithin the specific Examples.

The size exclusion chromatography (SEC) system used in the followingExamples is as follows: Column: Superdex Peptide 10/300 GL, GEHealthcare P/N 17-5176-01; Mobile Phase: 700 mL Milli-Q Plus water, 300mL acetonitrile; 1.0 mL trifluoroacetic acid; Flow Rate: 0.4 mL/minute;Temperature: ambient (˜21° C.); Detection: UV light at 214 nm, 280 nm;visible light at 476 nm; Injection: 10 μL; Run Time: 70 minutes; SamplePreparation: dilute 0.6-1.1 g to 10 mL with Mobile Phase; Calibration:by 6 purified reference proteins, 14.2-160 kD.

The high molecular weight lutein assembly is determined as follows: SECpeak eluting ≦20.0 minutes, using visible light detection at 476 nm (alutein absorbance maximum). The high MW lutein assembly is a waterdispersible or water soluble, protein-dominant, macromolecular aggregatein which lutein is bound. The size of the high MW lutein assembly liesapproximately in the 60-380 kD range (˜60-95 Å). The concentration ofthe high MW lutein assembly is quantified as peak area (mAU-min at 476nm) per mg of sample injected. The values reported as high MW luteinassembly are not absolute concentrations of bound lutein, but areinstead relative measures of the lutein bound in these high MW but waterdispersible/soluble, macromolecular aggregates.

The protein >137 kD is determined as follows: SEC peak eluting ≦18.0minutes, using UV detection at 214 nm (a peptide bond and amino acidside chain signal). The peak consists predominantly of aggregatedproteins; e.g., its size is larger than all major milk proteins (˜14 to25 kD), so that the protein in this peak (>137 kD) is regarded asaggregated but still water dispersible/soluble, and capable ofencapsulating lutein and other oil soluble vitamins/nutrients. It isquantified as g of protein per kg of sample. Peak area of the protein ismeasured versus a known concentration to provide the g/kg of saidprotein.

The bound lipophile index is determined as follows: a non-specificindicator of lipophilic nutrients (including lutein and any combinationof other oil soluble vitamins) in the high MW protein peak (describedabove). The value reported is the peak area ratio, 280 nm/214 nm, forthe SEC peak eluting ≦18.0 minutes, a relative marker of lipophilicnutrient concentration in the high MW aggregate. The validity of theindex is based on the fact that the 280 nm/214 nm ratios for lipophilicnutrients is >than that for proteins.

Examples 1-5

Examples 1-5 illustrate powdered infant formulas of the presentdisclosure, the ingredients of which are listed in the table below. Allingredient amounts are listed as kg per 1000 kg batch, unless otherwisespecified.

TABLE 3 Nutritional Composition Ingredients for Examples 1-5 and otherExemplified Compositions. INGREDIENT EXAMPLE 1 EXAMPLE 2 EXAMPLE 3EXAMPLE 4 EXAMPLE 5 Corn Syrup 504.1 504.1 504.1 504.1 504.1 Soy ProteinIsolate - 144.8 144.8 144.8 144.8 144.8 5% DH Sunflower Oil 112.5 112.5101.3 112.5 112.5 Sucrose 98.3 98.3 98.3 98.3 98.3 Soy Oil 83.9 83.975.5 83.9 83.9 Coconut Oil 75.6 75.6 68.0 75.6 75.6Fructooligosaccharides 17.0 17.0 17.0 17.0 17.0 Potassium Citrate 16.516.5 16.5 16.5 16.5 Calcium Phosphate 16.4 16.4 16.4 16.4 16.4 SodiumChloride 3.8 3.8 3.8 3.8 3.8 ARA Oil 3.0 3.0 3.0 3.0 3.0 MagnesiumChloride 2.8 2.8 2.8 2.8 2.8 L-Methionine 1.7 1.7 1.7 1.7 1.7 AscorbicAcid 1.1 1.1 1.1 1.1 1.1 DHA Oil 1.1 1.1 1.1 1.1 1.1 Mono-and approx.approx. approx. approx. approx. Diglyceride/Lutein 622.0 g 590.9 g 27.2653.1 g 685.0 g Blend Monodiglycerides and 621.0 g 590.3 g 27.0 652.7 g683.5 g Diglycerides Lutein 945.0 mg 566 mg 200 g 392 mg 1.5 g CholineChloride 507.7 g 507.7 g 507.7 g 507.7 g 507.7 g Taurine 457.5 g 457.5 g457.5 g 457.5 g 457.5 g Inositol 353.0 g 353.0 g 353.0 g 353.0 g 353.0 gAscorbyl Palmitate 347.5 g 347.5 g 347.5 g 347.5 g 347.5 g FerrousSulfate 319.2 g 319.2 g 319.2 g 319.2 g 319.2 g Mixed Tocopherols 157.2g 157.2 g 157.2 g 157.2 g 157.2 g L-Carnitine 112.7 g 112.7 g 112.7 g112.7 g 112.7 g Niacinamide 97.9 g 97.9 g 97.9 g 97.9 g 97.9 gd-alpha-tocopheryl 78.8 g 78.8 g 78.8 g 78.8 g 78.8 g acetate d-Calcium58.7 g 58.7 g 58.7 g 58.7 g 58.7 g Pantothenate Zinc 56.0 g 56.0 g 56.0g 56.0 g 56.0 g Iron 16.9 g 16.9 g 16.9 g 16.9 g 16.9 g Thiamine 15.2 g15.2 g 15.2 g 15.2 g 15.2 g Vitamin A palmitate 14.8 g 14.8 g 14.8 g14.8 g 14.8 g Copper 7.2 g 7.2 g 7.2 g 7.2 g 7.2 g Riboflavin 6.7 g 6.7g 6.7 g 6.7 g 6.7 g Pyridoxine 6.1 g 6.1 g 6.1 g 6.1 g 6.1 gHydrochloride Folic Acid 2.1 g 2.1 g 2.1 g 2.1 g 2.1 g Potassium Iodide1.1 g 1.1 g 1.1 g 1.1 g 1.1 g Phylloquinone 857.1 mg 857.1 mg 857.1 mg857.1 mg 857.1 mg Vitamin D3 103.7 mg 103.7 mg 103.7 mg 103.7 mg 103.7mg Lycopene 980.0 mg 980.0 mg 980.0 mg 980.0 mg 980.0 mg Biotin 592.5 mg592.5 mg 592.5 mg 592.5 mg 592.5 mg Beta-Carotene 215.6 mg 215.6 mg215.6 mg 215.6 mg 215.6 mg Selenium 147.0 mg 147.0 mg 147.0 mg 147.0 mg147.0 mg Cyanocobalamin 71.3 mg 71.3 mg 71.3 mg 71.3 mg 71.3 mg

Examples 6-10

Examples 6-10 illustrate powdered infant formulas of the presentdisclosure, the ingredients of which are listed in the table below. Allingredient amounts are listed as kg per 1000 kg batch, unless otherwisespecified.

TABLE 4 Nutritional Composition Ingredients for Examples 6-10 and otherExemplified Compositions. EXAMPLE INGREDIENT EXAMPLE 6 EXAMPLE 7 EXAMPLE8 EXAMPLE 9 10 Skim milk 2057.3 2057.3 2057.3 2057.3 2057.3 Lactose414.1 414.1 414.1 414.1 414.1 Sunflower oil 111.1 88.9 111.1 111.1 111.1Soy oil 83.3 66.6 83.3 83.3 83.3 Coconut oil 74.6 59.7 74.6 74.6 74.6GOS 72.3 72.3 72.3 72.3 72.3 Whey Protein 49.4 49.4 49.4 49.4 49.4Concentrate Whey protein 5.6 5.6 5.6 5.6 5.6 hydrolysate (<25DH)Potassium citrate 3.4 3.4 3.4 3.4 3.4 ARA 2.9 2.9 2.9 2.9 2.9 Calciumcarbonate 2.6 2.6 2.6 2.6 2.6 Potassium 2.3 2.3 2.3 2.3 2.3 hydroxideSoy lecithin 1.7 1.7 1.7 1.7 1.7 DHA 1.6 1.6 1.6 1.6 1.6 Ascorbic acid1.5 1.5 1.5 1.5 1.5 Choline bitartrate 1.2 1.2 1.2 1.2 1.2 Magnesium 1.21.2 1.2 1.2 1.2 chloride Mono-and approx. approx. approx. approx.approx. diglyceride/Lutein 1.10 53.8 0.99 1.155 1.21 blendMonoglycerides 1.10 53.5 0.99 1.153 1.21 and Diglycerides Lutein 2.65 g320 g 568 mg 2.3 g 385 mg Sodium chloride 1 1 1 1 1 Taurine 428 g 428 g428 g 428 g 428 g L-Tryptophan 277 g 277 g 277 g 277 g 277 g Cytidine5′- 257 g 257 g 257 g 257 g 257 g monophosphate Myo-inositol 257 g 257 g257 g 257 g 257 g Ascorbyl palmitate 200 g 200 g 200 g 200 g 200 gDisodium 187 g 187 g 187 g 187 g 187 g guanosine 5′- monophosphateDisodium uridine 163 g 163 g 163 g 163 g 163 g 5′-monophosphate Ferroussulfate 130.6 g 130.6 g 130.6 g 130.6 g 130.6 g monohydrate RRR-α- 119.7g 119.7 g 119.7 g 119.7 g 119.7 g Tocopheryl acetate Tocopherol-2 106.7g 106.7 g 106.7 g 106.7 g 106.7 g Zinc sulfate 104 g 104 g 104 g 104 g104 g monohydrate Adenosine 5′- 98 g 98 g 98 g 98 g 98 g monophosphateFerrous sulfate 88 g 88 g 88 g 88 g 88 g Mixed tocopherols 77 g 77 g 77g 77 g 77 g Niacinamide 59 g 59 g 59 g 59 g 59 g L-Carnitine 43 g 43 g43 g 43 g 43 g Calcium d- 42 g 42 g 42 g 42 g 42 g pantothenate Retinylpalmitate 13 g 13 g 13 g 13 g 13 g Copper sulfate 11 g 11 g 11 g 11 g 11g Thiamin HCl 9 g 9 g 9 g 9 g 9 g Pyridoxine HCl 5 g 5 g 5 g 5 g 5 gRiboflavin 5 g 5 g 5 g 5 g 5 g Manganese sulfate 3 g 3 g 3 g 3 g 3 gmonohydrate Folic acid 796 mg 796 mg 796 mg 796 mg 796 mg β-Carotene 770mg 770 mg 770 mg 770 mg 770 mg Phylloquinone 618 mg 618 mg 618 mg 618 mg618 mg Potassium iodide 550 mg 550 mg 550 mg 550 mg 550 mg d-Biotin 238mg 238 mg 238 mg 238 mg 238 mg Sodium selenate 117 mg 117 mg 117 mg 117mg 117 mg Cholecalciferol 93 mg 93 mg 93 mg 93 mg 93 mg Cyanocobalamin 8mg 8 mg 8 mg 8 mg 8 mg

Examples 11-15

Examples 11-15 illustrate powdered adult formulas of the presentdisclosure, the ingredients of which are listed in the table below. Allingredient amounts are listed as kg per 1000 kg batch, unless otherwisespecified.

TABLE 5 Nutritional Composition Ingredients for Examples 11-15 and otherExemplified Compositions. EXAMPLE EXAMPLE EXAMPLE EXAMPLE EXAMPLEINGREDIENT 11 12 13 14 15 Corn syrup 259.86 259.86 259.86 259.86 259.86Corn maltodextrin 216.25 216.25 216.25 216.25 216.25 Sucrose 177.95177.95 177.95 177.95 177.95 Corn oil 155.4 155.4 155.4 155.4 155.4Sodium and 152.43 152.43 152.43 152.43 152.43 calcium caseinates AcidCasein 132.97 132.97 132.97 132.97 132.97 Calcium 19.46 19.46 19.4619.46 19.46 Caseinate Soy protein isolate 28.94 28.94 28.94 28.94 28.94Mono-and approx. approx. approx. 2.3 approx. 7.8 approx.diglyceride/Lutein 15.53 15.6 5.91 Blend Monoglycerides 15.50 15.57 2.37.8 5.91 and diglycerides Lutein 2.9 g 34.0 g 5.0 g 2.5 g 12.5 gArtificial flavoring 14.55 14.55 14.55 14.55 14.55 Potassium citrate10.75 10.75 10.75 10.75 10.75 20% 3.22 3.22 3.22 3.22 3.22 PotassiumCitrate Potassium 7.54 7.54 7.54 7.54 7.54 Citrate Magnesium 7.53 7.537.53 7.53 7.53 chloride Calcium phosphate 6.23 6.23 6.23 6.23 6.23Sodium citrate 5.66 5.66 5.66 5.66 5.66 Potassium chloride 3.79 3.793.79 3.79 3.79 Soy lecithin 3.44 3.44 3.44 3.44 3.44 Ascorbic acid 1.911.91 1.91 1.91 1.91 Choline chloride 1.64 1.64 1.64 1.64 1.64 Zincsulfate 155.8 g 155.8 g 155.8 g 155.8 g 155.8 g dl-alpha-tocopheryl129.6 g 129.6 g 129.6 g 129.6 g 129.6 g acetate Niacinamide 124.2 g124.2 g 124.2 g 124.2 g 124.2 g Ferrous sulfate 121.7 g 121.7 g 121.7 g121.7 g 121.7 g Calcium 80.3 g 80.3 g 80.3 g 80.3 g 80.3 g pantothenateManganese sulfate 38.7 g 38.7 g 38.7 g 38.7 g 38.7 g Cupric sulfate 21.1g 21.1 g 21.1 g 21.1 g 21.1 g Thiamine chloride 20.5 g 20.5 g 20.5 g20.5 g 20.5 g hydrochloride Pyridoxine 19.8 g 19.8 g 19.8 g 19.8 g 19.8g hydrochloride Riboflavin 16.0 g 16.0 g 16.0 g 16.0 g 16.0 g Vitamin A10.4 g 10.4 g 10.4 g 10.4 g 10.4 g palmitate Folic acid 2.8 g 2.8 g 2.8g 2.8 g 2.8 g Biotin 2.4 g 2.4 g 2.4 g 2.4 g 2.4 g Chromium chloride 1.3g 1.3 g 1.3 g 1.3 g 1.3 g Sodium molybdate 1.0 g 1.0 g 1.0 g 1.0 g 1.0 gPotassium iodide 0.44 g 0.44 g 0.44 g 0.44 g 0.44 g Sodium selenate 0.43g 0.43 g 0.43 g 0.43 g 0.43 g Phylloquinone 0.22 g 0.22 g 0.22 g 0.22 g0.22 g Cyanocobalamin 0.05 g 0.05 g 0.05 g 0.05 g 0.05 g Vitamin D3 0.03g 0.03 g 0.03 g 0.03 g 0.03 g

Examples 16-18

Examples 16-18 illustrate liquid adult formulas of the presentdisclosure, the ingredients of which are listed in the table below. Allingredient amounts are listed as kg per 1000 kg batch, unless otherwisespecified.

TABLE 6 Nutritional Composition Ingredients for Examples 16-18 and otherExemplified Compositions. Example Example Example Ingredient 16 17 18Water QS QS QS Corn Syrup 33 kg 33 kg 33 kg Maltodextrin 28 kg 28 kg 28kg Sucrose 19.4 kg 19.4 kg 19.4 kg Mono- and 18.3 kg 6.1 kg 2.4 kgDiglyceride/DHA premix Monoglycerides 9.6 kg 5.1 kg 2 kg andDiglycerides Docosahexaenoic 8.7 kg 1 kg 0.4 kg Acid Oil (60% DHA invegetable oil) Caseinate 8.7 kg 8.7 kg 8.7 kg High Oleic — 2 kg 3.3 kgSafflower Oil Canola Oil — 2 kg 3.3 kg Soy Protein 3.7 kg 3.7 kg 3.7 kgWhey Protein 3.2 kg 3.2 kg 3.2 kg Caseinate 2.9 kg 2.9 kg 2.9 kg CornOil — 1 kg 1.6 kg Tricalcium 1.4 kg 1.4 kg 1.4 kg Phosphate PotassiumCitrate 1.3 kg 1.3 kg 1.3 kg Magnesium 952 gm 952 gm 952 gm PhosphateLecithin 658 gm 658 gm 658 gm Magnesium 558 gm 558 gm 558 gm chlorideVanilla Flavor 544 gm 544 gm 544 gm Sodium Chloride 272 gm 272 gm 272 gmCarrageenan 227 gm 227 gm 227 gm Choline chloride 218 gm 218 gm 218 gmUTM/TM Premix 165 gm 165 gm 165 gm Potassium Chloride 146 gm 146 gm 146gm Ascorbic Acid 145 gm 145 gm 145 gm Sodium Citrate 119 gm 119 gm 119gm Potassium 104 gm 104 gm 104 gm Hydroxide Docosahexaenoic 8.7 kg 1 kg1 kg Acid Oil (60% DHA in vegetable oil) WSV Premix 33 gm 33 gm 33 gmVit DEK Premix 29 gm 29 gm 29 gm Vitamin A 3.7 gm 3.7 gm 3.7 gmPotassium Iodide 86 mcg 86 mcg 86 mcg

Example 19

In this Example, the effect of mixing lutein, a polar lipid nutrient, ina combination of monoglycerides and diglycerides on the luteinbioavailability is evaluated and compared to the bioavailability oflutein that is mixed in typical triglyceride-based oils.

Male Sprague Dawley rats weighing between 280 and 330 grams were all feda normal commercially available Purina Rat Chow for one week. Rats werethen fasted overnight prior to surgery, and under anesthesia, alaparotomy was performed, and the intestinal lymph duct was cannulatedaccording to the procedure of Tso et al., “The Absorption of Lipid andLipoprotein Synthesis,” Lipid Research Methodology, Chapter 5: 191-216(1984) Alan R. Liss, Inc., NY, N.Y., hereby incorporated by reference tothe extent consistent herewith. The superior mesenteric artery wasisolated, but not occluded. A silicon infusion tube (1.6 mm OD) wasplaced in the stomach for future infusion of a test sample. The fundicincision was closed by a purse string suture. The rats were allowed torecover for 24 hours before infusion began.

The test samples included the following solutions: (1) lutein insafflower oil (control); (2) lutein in monoglycerides and diglyceridesderived from corn oil; and (3) lutein in monoglycerides and diglyceridesderived from high oleic acid sunflower oil.

The rats were randomly assigned to three groups. The animals wereintragastrically infused 24 hours after surgery with 20 mg/kg luteinfrom their respective lutein-containing solutions.

Lymph was collected in pre-cooled tubes 1 hour before the luteininfusions (fasting) and then hourly for 6 hours after initiation ofinfusion. At the end of the 6 hours infusion, the rats were sacrificedby exsanguination.

The lymph lipids were extracted and analyzed for lutein concentrationusing high-performance liquid chromatography with programmed wavelengthultraviolet detection (Craft Technologies, Wilson, N.C.).

FIG. 1 shows the hourly lymph flow rate over a period of 6 hoursfollowing lutein administration. FIG. 2 shows the hourly lutein outputin lymph over a 6 hour period after lutein administration. FIG. 3 showscumulative lymphatic lutein absorption over a 6 hour period after luteinadministration. FIG. 4 shows the percent change in lymphatic luteinabsorption of lutein with mono- and diglycerides versus control over aperiod of 6 hours.

As shown in FIG. 1, the mean fasting lymph flow for all three groups ofrats varied between 2.3 and 2.6 mL/h. In all groups, lymph flowincreased significantly after lutein infusion and reached a maximumoutput between 2.7 and 3.9 mL/h during hours 2-3 after lipid infusion.There were no differences in lymph flow rates between the groupsindicating that the use of lutein in mono- and diglycerides did notsignificantly affect lymph flow rates versus the lutein in triglyceridecontrol. FIG. 2 shows the lymphatic lutein output in μg/h during thefirst 6 hours after gastric feeding of the 3 different lutein samples.The lymphatic output of lutein increased in all groups during the first3 hours and reached a steady state from hour 3 to hour 5 after luteinadministration. However, there was a significant increase in the amountsof lymphatic lutein output 2-6 hours after lutein administration withlutein given with mono and diglycerides (corn and HOSO) as compared withlutein in the triglyceride control. FIG. 3 shows the above luteinabsorption improvements with mono and diglycerides by demonstrating asignificantly higher cumulative lutein absorption over the 6 hourabsorption period (area under the curve (AUC)) versus the lutein in thetriglyceride control. When this data is expressed as a percent change inlymphatic lutein absorption, as shown in FIG. 4, there was a 78%-88%increase in lymphatic lutein absorption of lutein with mono anddiglycerides versus the control over the 6 hour infusion period. Thisdata indicates that the polar lipid nutrient lutein has improvedbioavailability in a mixture of monoglycerides and diglycerides ascompared to triglyceride oils.

Example 20

In this Example, the effect of the addition of an activated premixcomprising lutein and the effect of the addition of a protected premixon the bioavailability of the lutein in a nutritional composition isevaluated and compared to the bioavailability of lutein that istypically added to triglyceride-based oils prior to addition to thenutritional composition.

Male Sprague Dawley rats weighing between 280 and 330 grams were all feda normal commercially available Purina Rat Chow for one week. Rats werethen fasted overnight prior to surgery, and under anesthesia, alaparotomy was performed, and the intestinal lymph duct was cannulatedaccording to the procedure of Tso et al., “The Absorption of Lipid andLipoprotein Synthesis,” Lipid Research Methodology, Chapter 5: 191-216(1984) Alan R. Liss, Inc., NY, N.Y., hereby incorporated by reference tothe extent consistent herewith. The superior mesenteric artery wasisolated, but not occluded. A silicon infusion tube (1.6 mm OD) wasplaced in the stomach for future infusion of a test sample. The fundicincision was closed by a purse string suture. The rats were allowed torecover for 24 hours before infusion began.

The test samples included the following nutritional compositions:(AET-C) lutein added to high oleic safflower oil at 120° F. for 10minutes, added to the protein-in-fat slurry (control); (AET-1) luteinpremixed with monoglycerides and diglycerides at 120° F. for 10 minutes,lecithin added to the premix post heating and allowed to mix for 5minutes, added to the protein-in-fat slurry to form an MDG-protectedpremix prior to addition to the remainder of the composition; (AET-2)lutein premixed with monoglycerides and diglycerides at 120° F. for 10minutes, added to the protein-in-water slurry to form an activatedpremix, prior to addition to the remainder of the composition; (AET-3)lutein premixed with monoglycerides and diglycerides at 120° F. for 10minutes added to the protein-in-fat slurry prior to addition to theremainder of the composition, and (AET-4) lutein premixed withmonoglycerides and diglycerides at 120° F. for 10 minutes, choline-addedas choline chloride to the premix post heating and allowed to mix for 5minutes, added to the protein-in-water slurry to form an activatedpremix prior to addition to the remainder of the composition. Thecompositions for each of the test samples are provided in detail in thefollowing table.

TABLE 7 Nutritional Composition Ingredients for Example 20 and otherExemplified Compositions. AET-C AET-1 AET-2 AET-3 AET-4 Amount AmountAmount Amount Amount per per per per per Ingredient Name: 1000 kg Unit1000 kg Unit 1000 kg Unit 1000 kg Unit 1000 kg Unit Ingredient WaterQ.S. kg Q.S. kg Q.S. kg Q.S. kg Q.S. kg Lactose 49.27 kg 49.27 kg 49.27kg 49.27 kg 49.27 kg Non-Fat Dry Milk 22.06 kg 22.06 kg 22.06 kg 22.06kg 22.06 kg High Oleic Safflower 13.44 kg 12.78 kg 12.78 kg 12.78 kg12.78 kg Oil Soy Oil 10.06 kg 9.57 kg 9.57 kg 9.57 kg 9.57 kg CoconutOil 9.60 kg 9.13 kg 9.13 kg 9.13 kg 9.13 kg Galacto-oligosaccharides9.40 kg 9.40 kg 9.40 kg 9.40 kg 9.40 kg Whey Protein 6.31 kg 6.31 kg6.31 kg 6.31 kg 6.31 kg Concentrate Monoglyceride and 1.62 kg 1.62 kg1.62 kg 1.62 kg Diglyceride Oil Ascorbic Acid 611.99 g 611.99 g 611.99 g611.99 g 611.99 g Calcium Carbonate 559.70 g 559.70 g 559.70 g 559.70 g559.70 g Potassium Citrate 545.40 g 545.40 g 545.40 g 545.40 g 545.40 gARASCO Mortierella 395.30 g 395.30 g 395.30 g 395.30 g 395.30 g AlpinaOil KOH 45% 368.00 g 368.00 g 368.00 g 368.00 g 368.00 g Centrol(Lecithin) 346.99 g 346.99 g 346.99 g 346.99 g 346.99 g Myverol 346.99 g346.99 g 346.99 g 346.99 g 346.99 g Nucleotide-Choline 314.15 g 314.15 g314.15 g 314.15 g 314.15 g Premix DHASCO 208.80 g 208.80 g 208.80 g208.80 g 208.80 g Crypthecodinium Cohnii Oil Vit/Min/Taur Premix 176.00g 176.00 g 176.00 g 176.00 g 176.00 g Seakem RLC 175.26 g 175.26 g175.26 g 175.26 g 175.26 g Carrageenan Sodium Citrate 163.00 g 163.00 g163.00 g 163.00 g 163.00 g Sodium Chloride 145.30 g 145.30 g 145.30 g145.30 g 145.30 g Magnesium Chloride 126.50 g 126.50 g 126.50 g 126.50 g126.50 g Potassium Phosphate 117.60 g 117.60 g 117.60 g 117.60 g 117.60g Dibasic Ferrous sulfate 66.50 g 66.50 g 66.50 g 66.50 g 66.50 gCholine Chloride 44.97 g 44.97 g 44.97 g 44.97 g 44.97 g Vitamin A, D3,E, K1 35.80 g 35.80 g 35.80 g 35.80 g 35.80 g Magnesium Sulfate 27.40 g27.40 g 27.40 g 27.40 g 27.40 g Zinc Sulfate 17.90 g 17.90 g 17.90 g17.90 g 17.90 g Lutein 12.35 g 12.35 g 12.35 g 12.35 g 12.35 gL-Carnitine 5.50 g 5.50 g 5.50 g 5.50 g 5.50 g Cupric Sulfate 1.90 g1.90 g 1.90 g 1.90 g 1.90 g Manganese Sulfate 110.00 mg 110.00 mg 110.00mg 110.00 mg 110.00 mg Citric Acid (Processing As As As As As aid)needed needed needed needed needed Potassium Iodide As As As As Asneeded needed needed needed needed Sodium Selenate As As As As As neededneeded needed needed needed Potassium Chloride As As As As As neededneeded needed needed needed

The rats were randomly assigned to five groups. The animals wereintragastrically infused 24 hours after surgery with 3 ml of nutritionalformula that delivered 5.8 μg lutein per animal from their respectivelutein-containing solutions.

Lymph was collected in pre-cooled tubes 1 hour before the nutritionalcomposition infusions (fasting) and then hourly for 8 hours afterinitiation of infusion. At the end of the 8 hours infusion, the ratswere sacrificed by exsanguination.

The lymph lipids were extracted and analyzed for lutein concentrationusing high-performance liquid chromatography with programmed wavelengthultraviolet detection (Craft Technologies, Wilson, N.C.).

FIG. 5 shows the hourly lymph flow rate over a period of 8 hoursfollowing administration of liquid infant compositions containinglutein. FIG. 6 shows the hourly lutein output in lymph over an 8 hourperiod after administration of liquid infant compositions containinglutein. FIG. 7 shows cumulative lymphatic lutein absorption over a 6hour and 8 hour period after administration of liquid infantcompositions containing lutein. FIG. 8 shows the percent change inlymphatic lutein absorption over a period of 6 hours and 8 hoursfollowing administration of liquid infant compositions containing luteinversus control.

As shown in FIG. 5, the mean fasting lymph flow for all three groups ofrats varied between 2.4 and 2.7 mL/h. In all groups, lymph flowincreased significantly after nutritional composition infusion andreached a maximum output between 2.8 and 3.1 mL/h during hours 2-3 afternutritional composition infusion. There were no differences in lymphflow rates between the groups indicating that the use of a nutritionalcomposition including either an activated premix with lutein or anMDG-protected premix with lutein, did not significantly affect lymphflow rates versus the control nutritional composition including luteinin triglyceride. FIG. 6 shows the lymphatic lutein output in μg/h duringthe first 8 hours after gastric feeding of the 5 different nutritionalcomposition samples. The lymphatic output of lutein increased in allgroups during the first 3 hours and reached a steady state from hour 2to hour 3 after nutritional formula administration. However, there was asignificant increase in the amounts of lymphatic lutein output 2-8 hoursafter lutein administration when lutein was given in the form of anactivated premix (AET-2, AET-4) or with lutein given in the form of anMDG-protected premix (AET-1), as compared with lutein mixed directlywith the triglycerides control or non-activated premix containinglutein, monoglycerides and diglycerides (AET-C and AET-3, respectively).FIG. 7 shows the lutein absorption improvements with an activated premixwith lutein or an MDG-protected premix with lutein by demonstratingsignificantly higher cumulative lutein absorption over the 6 and 8 hourabsorption period (area under the curve (AUC)) versus the lutein in thecontrol and non-activated samples. When this data is expressed as apercent change in lymphatic lutein absorption, as shown in FIG. 8, therewas a 106%-170% increase in lymphatic lutein absorption of compositionscomprising activated premix with lutein or an MDG-protected premix withlutein versus the control and non-activated sample over the 6 hourinfusion period. There was a 121%-165% increase in lymphatic luteinabsorption of the compositions comprising an activated premix withlutein or an MDG-protected premix with lutein versus the control andnon-activated sample over the 8 hour infusion period. These dataindicate that the lipophilic compound lutein has improvedbioavailability when in an activated premix or an MDG-protected premixas compared to when it is in triglyceride oils.

Example 21

In this Example, the test samples evaluated in Example 20 are furtherevaluated for lutein that is not solubilized by the fat in the samplenutritional compositions. The five samples, AET-C, AET-1, AET-2, AET-3and AET-4 were each subjected to high speed centrifugation (31,000×g;20° C.; 4 h). The visible absorbance of the resulting aqueous fractions(after 2:8 dilution with water, followed by syringe filtration through a0.45 μm PTFE membrane) was measured at 476 nm (a lutein absorbancemaximum). The absorbance readings (expressed as milliabsorbance unitsper gram of aqueous fraction (mAU/g)) are reported in the table below,along with a ranking of the in vivo lutein response for the samevariations tested in Example 20.

TABLE 8 Relationship between Lutein in the Aqueous Fraction with in vivoResponse Visible light absorbance, In vivo ranking of lutein Sample IDmAU/g, 476 nm response AET-C 57.6 1 (lowest response) AET-1 62.7 4 AET-261.3 3 AET-3 59.4 2 AET-4 64.9 5 (highest response)

As may be seen from the table, the absorbance values are correlated withthe in vivo findings in that each of the sample nutritional compositionscomprising an activated premix (AET-2 and AET-4) or an MDG-protectedpremix (AET-1) have higher lutein absorbance values than those samplenutritional compositions that do not comprise an activated premix (AET-Cand AET-3). This means that more lutein is present in the aqueousfraction of the sample nutritional compositions comprising an activatedpremix or an MDG-protected premix. Thus, at least in relation to thesample nutritional compositions, the bioavailability of lutein wasaccurately predicted in vitro utilizing this method.

In this Example, the effect of the addition of an activated premix or anMDG-protected premix to a nutritional composition on the bioavailabilityof other lipophilic compounds such as total triglycerides,phospholipids, arachidonic acid (ARA) and docosahexaenoic acid (DHA) isevaluated and compared to the bioavailability of the lipophiliccompounds that are consumed in the absence of the activated premix orthe MDG-protected premix.

A similar protocol for animal infusions and lymph collection asdescribed in Example 20 was followed, including the use of the same testsamples. The lymph lipids were extracted and analyzed for triglycerides,phospholipids, ARA and DHA concentration using high-performance liquidchromatography.

FIG. 9 shows the hourly lymphatic triglyceride output over a period of 6hours following nutritional composition administration. FIG. 10 showsthe hourly lymphatic phospholipid output over 6 hours followingnutritional composition administration.

FIG. 11 and FIG. 12 show the change in lymphatic absorption of ARA andDHA from fasting, respectively, over a period of 6 hours followingnutritional composition administration.

As shown in FIG. 9, the lymphatic output of triglycerides increased inall groups during the 6 hours post administration of nutritionalcomposition samples. However, there was a significant increase in theamounts of lymphatic triglyceride output 2-6 hours after sampleadministration with an activated premix (AET-2, AET-4) or anMDG-protected premix (AET-1) as compared with the samples having themonoglycerides and diglycerides mixed directly with the triglycerides ina non-activated premix or a non-MDG-protected premix (AET-C and AET-3).Similarly, FIG. 10 shows that the lymphatic output of phospholipidsincreased in all groups during the 6 hours post sample administration.There was a significant increase in the amount of lymphatic phospholipidoutput 2-6 hours after sample administration with a given in anactivated premix (AET-2, AET-4) or an MDG-protected premix (AET-1) ascompared with the samples having the monoglycerides and diglyceridesmixed directly with the triglycerides in a non-activated premix or anon-MDG-protected premix (AET-C and AET-3).

As shown in FIG. 11, there was a significant increase in lymph levels ofARA from fasting during the 6 hours post administration of thenutritional compositions containing an activated premix (AET-2) or anMDG-protected premix (AET-1) as compared with the non-activated ornon-MDG-protected control. FIG. 12 shows similar results bydemonstrating a significant increase in lymph levels of DHA from fastingduring the 6 hours post administration of the nutritional formulascontaining an activated premix (AET-2) or an MDG-protected premix ascompared with the non-activated or non-MDG-protected control.

Example 22

A study was conducted to compare a commercially representativenutritional powder (details of powder components are listed below) withand without MDG. The nutritional powder batches (control and MDG) werereconstituted (152 g/L), centrifuged (31,000×g/20° C./4 h), and analyzedas outlined below. The metric used to analyze the bound lutein is theearly eluting peak of the SEC chromatogram, which corresponds to thehigh molecular weight peak. The numerical value of this peak is the areaof the peak at 476 nm per microliter of sample injected. Additionally,the cream layer, which is the upper layer obtained upon the high speedcentrifugation, is used to examine the mechanism of action for enhancedlutein solubility, as well as to determine which proteins areresponsible for the bound lutein assemblies.

The markers of bound lutein (high MW size exclusion chromatography (SEC)peak area at 476 nm) were significantly higher for MDG batches versuscontrol. The cream layer protein comparison and the attachedchromatograms were consistent with MDG-facilitated formation of high MW,but water dispersible (aqueous fraction), and even water soluble (100 kultrafiltrate), assemblies of native proteins (casein, α-lactalbumin,β-lactoglobulin) and lutein.

TABLE 9 Nutritional Composition Ingredients for Example 22 and otherExemplified Compositions. INGREDIENTS (MDG INGREDIENTS (Control AMOUNTPER 1000 Incorporated Formula) AMOUNT PER 1000 KG Formula) KG Non FatDry Milk 442 kg Non Fat Dry Milk 442 kg Lactose 167.8 kg Lactose 167.8kg Sucrose 111.9 kg Sucrose 111.9 kg High oleic safflower oil 86.6 kgHigh oleic safflower oil 89.9 kg Soy oil 67.4 kg Soy oil 67.4 kg CoconutOil 64.4 kg Coconut Oil 64.4 kg Galactooligosaccharides (GOS) 36.2 kgGalactooligosaccharides (GOS) 36.2 kg Whey protein concentrate (75%)35.7 kg Whey protein concentrate (75%) 35.7 kg Potassium Citrate 6.8 kgPotassium Citrate 6.8 kg Capmul GM040 (MDG Oil) 3.29 kg CalciumCarbonate 2.7 kg Calcium Carbonate 2.7 kg Sodium Chloride 1.7 kg SodiumChloride 1.7 kg Nucleotide/Choline Premix 1.6 kg Nucleotide/CholinePremix 1.6 kg Choline bitartrate 805.5 g Choline bitartrate 805.5 gCytidine 5′-monophosphate 166.3 g Cytidine 5′-monophosphate 166.3 gDisodium guanosine 121 g 5′-monophosphate Disodium guanosine 5′- 121 gDisodium uridine 5′- 105.4 g monophosphate monophosphate Disodiumuridine 5′- 105.4 g Adenosine 5′- 63.2 g monophosphate monophosphateAdenosine 5′-monophosphate 63.2 g Maltodextrin (premix diluent) q.s.Maltodextrin (premix diluent) q.s. Lecithin, PCR negative, IP 1.4 kgLecithin, PCR negative, IP 1.4 kg Ascorbic Acid 1.4 kg Ascorbic Acid 1.4kg Tricalcium phosphate 1.16 kg Tricalcium phosphate 1.16 kg WaterSoluble Vitamin 869.7 g and Mineral Premix Water Soluble Vitamin andMineral 869.7 g Taurine 265.8 g Premix Taurine 265.8 g m-Inositol 193.0g m-Inositol 193.0 g Zinc Sulfate, Monohydrate 89.0 g Zinc Sulfate,Monohydrate 89.0 g Niacinamide 56.7 g Niacinamide 56.7 g d-CalciumPantothenate 34.0 g d-Calcium Pantothenate 34.0 g Ferrous Sulfate, Dried29.8 g (Monohydrate) Ferrous Sulfate, Dried 29.8 g Copper Sulfate,Anhydrous 10.4 g (Monohydrate) Copper Sulfate, Anhydrous 10.4 g ThiamineChloride 8.80 g Hydrochloride Thiamine Chloride 8.80 g Riboflavin 3.88 gHydrochloride Riboflavin 3.88 g Pyridoxine Hydrochloride 3.56 gPyridoxine Hydrochloride 3.56 g Folic Acid 1.31 g Folic Acid 1.31 gManganese Sulfate, Monohydrate 1.01 g Manganese Sulfate, Monohydrate1.01 g Biotin 0.343 g Biotin 0.343 g Sodium Selenate, Anhydrous 0.206 gSodium Selenate, Anhydrous 0.206 g Cyanocobalamin 0.027 g Cyanocobalamin0.027 g Corn Starch (modified) 27.4 g (0.1% B12 spray dry) Corn Starch(modified) (0.1% 27.4 g Dextrose q.s. B12 spray dry) Dextrose q.s.Docosahexaenoic acid (DHA) 759.2 g Docosahexaenoic acid (DHA) 759.2 gVitamin A, D3, E, K1 premix 386.9 g Vitamin A, D3, E, K1 premix 386.9 gd-α-tocopheryl acetate 78.2 g d-α-tocopheryl acetate 78.2 g Vitamin Apalmitate 14.6 g Vitamin A palmitate 14.6 g Phylloquinone 851.2 mgPhylloquinone 851.2 mg Vitamin D3 103.0 mg Vitamin D3 103.0 mg Vegetableoil (premix diluent) q.s. Vegetable oil (premix diluent) q.s. CholineChloride 284 g Choline Chloride 284 g Magnesium Chloride 261.1 gMagnesium Chloride 261.1 g Ferrous Sulfate 222.0 g Ferrous Sulfate 222.0g Arachidonic acid 205.2 g Arachidonic acid 205.2 g Ascorbyl Palmitate164.4 g Ascorbyl Palmitate 164.4 g Tocopherol-2 food grade antioxidant87.8 g Tocopherol-2 food grade antioxidant 87.8 g Mixed tocopherols 63.7g Mixed tocopherols 63.7 g Vegetable oil (diluent) q.s. Vegetable oil(diluent) q.s. Lutein 20% 12.5 g Lutein 20% 12.5 g Lutein 2.5 g Lutein2.5 g Beta-Carotene (30%) 3.22 g Beta-Carotene (30%) 3.22 g β-carotene966 mg β-carotene 966 mg Potassium Hyrdoxide as needed PotassiumHyrdoxide as needed

Example 23

A study was conducted to compare the bound lutein size estimates withinthe aqueous fraction. Bound lutein size estimates were investigated viaSAXS, SEC, and MALDI-TOF.

In summary, the bound lutein may be placed into two or three categories:(a) large (˜100-1000 Å, or >250 kD) water soluble/dispersible aggregatesthat are not discrete structures and span a broad size range asdetermined by SAXS and SEC analysis; (b) native protein/lutein complexes(˜10-50 kD, ˜27 TO 57 Å) that are generally discrete, well-definedstructures, which is supported by MALDI-TOF and SEC data; and (c) arelatively small (˜8 Å, or ˜3-4 kD) crystal. The small structure may ormay not comprise lutein.

Example 24

A study was conducted to examine the grand average of hydropathicityindex (GRAVY) of milk proteins. The GRAVY value is a measure ofhydrophobicity and water solubility, and may be predictive of aprotein's ability to bind a lipophilic molecule such as lutein.

The lutein binding capacities of eight major milk proteins werepredicted from their GRAVY values using the plot listed below. The GRAVYvalues of said milk proteins are listed in the table below, as well astheir predicated lutein binding capacity. It was found that the milkproteins γ₂-casein, β-lactoglobulin, β-casein and α-lactalbumin had thehighest predicted lutein binding capacity of <2 μM, which correlateswith MALDI-TOF data implicating these proteins as binding participantsin the lutein/protein complexes. Accordingly, the GRAVY values were ableto correlate accurate lutein binding trends of proteins that have beenshown to efficiently interact with lutein within the nutritionalproduct. This results in GRAVY values potentially providing insight intoother potential proteins, and may be useful in optimizing thenutritional product system.

TABLE 10 GRAVY Value Analysis of Different Proteins Protein MW, kD GRAVYLutein K_(d), μM K_(d) Source StARD3 50.5 0.001 0.45 Published GSTP123.2 −0.131 1.35 Published HSA 66.5 −0.395 1.69 Published Milk proteinsγ₂-casein 11.8 −0.136 1.05 Predicted β-lactoglobulin 18.3 −0.162 1.13Predicted β-casein 23.6 −0.355 1.68 Predicted α-lactalbumin 14.2 −0.4531.96 Predicted BSA 66.4 −0.475 2.02 Predicted κ-casein 19.0 −0.557 2.26Predicted α-s₁-casein 23.0 −0.704 2.68 Predicted α-s2-casein 24.3 −0.9183.29 Predicted

Example 25

A study was conducted to investigate the size/mass of the high molecularweight lutein assemblies. A SEC calibration curve was determined toanalyze the aforementioned high molecular weight assemblies. Thesevalues were also compared to the data presented in Example 3, regardingsize of the high molecular weight lutein assembly.

The reference proteins, their SEC elution times and molecular weight arelisted in the table below, and were used to estimate the size of thehigh molecular weight lutein assembly. The SEC analysis was performed asrecited in the preceding Examples. By extrapolation from a referenceprotein plot, the size of the high molecular weight lutein assembly wasfound to be in the ranges of 60-380 kD, with a peak at 160 kD. Thesevalues correspond to particle size diameters of 60-96 Å, with a peak at78 Å. The estimated values presented below, appear to be consistent withSAXS data presented in Example 23, wherein it corresponds with theintermediated size category of 10-100 Å. It should be noted thatparticles in the large category, presented in Example 23, would beexcluded from the ultrafiltrate, and there may be larger luteinassemblies in the aqueous fraction prior to ultrafiltration.Additionally, it is noted that the 214 nm/280 nm ratio of the highmolecular weight lutein assembly was in the range of the referenceproteins, suggesting a protein presence in the high molecular weightassembly.

TABLE 11 Calibration of Assembly Size with Known Protein MolecularWeights Reference Protein SEC Elution Time, 214 nm/280 nm (Sigma,bovine) MW, kD minutes peak area ratio α-Lactalbumin 14.2 23.67 8.12β-Lactoglobulin 18.3 22.41 13.6 β-Casein 23.6 21.17 19.1 BSA 66.4 19.0117.3 Lactoferrin 76.1 19.08 10.6 IgG ~160 19.05 10.7 High MW luteinassembly Range 60 to 380 kD 15.7-20.0 10.6 (~60 to 95 Å) Peak 160 kD17.66 N/A (~78 Å)

Example 26

A study was conducted to examine the high molecular weight luteinassemblies in commercially available nutritional powder batches, bothbefore and after centrifugation. The nutritional powder batches (controland MDG incorporated) were reconstituted (152 g/L), and were analyzedbefore and after centrifuged (31,000×g/20° C./4 h). The components ofthese powders are listed in the table of Example 22. The table belowdisplays the high molecular weight lutein assembly analysis via SEC, andis represented as mAU-min/mg injected. The MDG composition shows higherinitial lutein presence in both the whole product and the aqueousfraction, wherein it shows even a higher difference in the aqueousfraction relative to the control composition.

It was found that the high molecular weight lutein assembly presentwithin the aqueous fraction was 22% higher than in the control batch.This indicates significantly enhanced solubility and/or stability in theMDG composition. Additionally, the chromatograms illustrate that somefraction (12.3% to 14.5%) of the high molecular weight lutein assemblyis recovered in the supernatant after high speed centrifugation, whichconveys that these assemblies are sufficiently soluble and stable thatthey were not removed or disassembled by the stresses associated withhigh speed centrifugation.

TABLE 12 Analysis of Commercially Representative Nutritional Compositioncomprising MDG Batch A545 (MDG Batch A544 (control) Incorporated) Wholeproduct 1977 2070 (w/o centrifugation) Aqueous Fraction 244 299 (aftercentrifugation) % Recovery 12.3% 14.5%

Example 27

A study was conducted to evaluate high molecular lutein assembliesfacilitated by the presence of MDG vs. high oleic safflower oil (HOSO).Bench scale model systems were prepared in nonfat dry milk (NFDM),wherein the NFDM was used at 10% (w/w), a lipid (HOSO or MDG), andlutein. Lutein was used at variable amounts, and these compositions werethen assessed for the formation of high molecular weight assemblies. Thedifferent compositions were centrifuged (31,000×g/20° C./1 h) to obtainaqueous fractions, which were compared by a pH 2.5 SEC as detailedabove, and lutein assemblies were analyzed by measuring a visible lightabsorbance at 476 nm, and by measuring visible light transmittance at860 nm Additionally, there was a group of MDG-batches that were notcentrifuged, so the whole product was analyzed for the presence oflutein assemblies.

The results demonstrate that HOSO has little capacity for facilitatingthe high molecular weight lutein assemblies at any of the differentlutein concentrations. In contrast, the MDG compositions exhibitedsignificant formation of high molecular weight lutein assemblies, whichmay be seen with the higher absorbance at 476 nm. Furthermore, thisexperiment revealed that lutein concentration greater than 1×, showed adecrease in lutein-assembly formation. This data suggests that there isan optimal ratio of lutein to MDG for the formation of high molecularweight lutein assemblies. Additionally, the non-centrifuged samplesreveal that approximately 36% of the total assembly is recoveredfollowing high-speed centrifugation.

Example 28

A study was conducted to analyze protein involvement with the highmolecular weight lutein assembly. These studies were performed onbatches that comprised MDG or HOSO. The different batch variations wereexamined using the same techniques as listed above. Specifically, benchscale model systems were prepared in nonfat dry milk, wherein the NFDMwas used at 10% (w/w), a lipid (HOSO or MDG), and lutein. Lutein wasused at variable amounts, and these compositions were then assessed forthe formation of high molecular weight assemblies. The differentcompositions were centrifuged (31,000×g/20° C./1 h) to obtain aqueousfractions, which were compared by a pH 2.5 SEC as detailed above.Protein was analyzed to examine the different molecular weight fractionsand their association with lutein.

The results demonstrated that MDG enhances the solubility and/or thestability of both molecular weight fractions, >57 kD and >137 kD. Thiswas especially enhanced in the 137 kD fraction for the MGD batches. Itshould be noted that the density of the protein assembly may be a factorin its centrifugation recovery, as would be expected as the lipidcontent of the assembly significantly increases, the assembly may bemore vulnerable to removal by high speed centrifugation. Additionally,as shown in the prior Example, lutein incorporation for compositionswith greater than 1× lutein actually decreased assembly formation. Thisfurther suggests that an optimal ratio/range of MDG and lutein areneeded.

TABLE 13 Model System Investigation of Assembly Formation High MW TubeLipid Lutein Centrifugation lutein assembly 1 HOSO 0X Yes 33.5 2 HOSO 1XYes 33.9 3 HOSO 2X Yes 37.2 4 HOSO 4X Yes 36.0 5 MDG 0X Yes 36.3 6 MDG1X Yes 50.0 7 MDG 2X Yes 42.2 8 MDG 4X Yes 41.6 5 MDG 0X No 101 6 MDG 1XNo 139 7 MDG 2X No 129 8 MDG 4X No 139

Example 29

A study was conducted to investigate the formation of high molecularweight lutein assemblies at different concentrations of MDG, HOSO andlutein. Specifically, lutein assembly was assessed by measuringmAU-min/mg at 476 nm, and by determining the presence of the >137 kDprotein fraction present in the aqueous fraction followingcentrifugation. These tests were performed as listed above, with thedifferent compositions being centrifuged (31,000×g/20° C./1 h) to obtainaqueous fractions, which were analyzed by SEC as detailed above.

In summary, the MDG batches outperformed the HOSO batches at both thepresence of lutein high molecular weight assemblies, as well as greateramounts of the >137 kD fraction being present within the aqueousfraction. Additionally, it should be noted that the size of the assemblyvaried, even though the MDG/lutein ratio was held constant. Since thelutein assemblies were increased using a combination of MDG and HOSO,their use within the composition may be advantageous.

TABLE 14 Model System Investigation II of Assembly Formation High MWlutein Aqueous assembly, Fraction mAU-min/mg, protein >137 kD, VariableHOSO, g/L MDG, g/L Lutein, μg/L 476 nm g/kg 1 0 6.67 10.4 59.7 0.517 2 04.44 6.94 58.0 0.498 3 0 2.22 3.47 40.8 0.447 4 6.67 0 10.4 38.1 0.412 54.44 0 6.94 37.4 0.408 6 2.22 0 3.47 35.2 0.380 7 3.33 3.33 10.4 55.80.490 8 1.11 1.11 3.47 45.7 0.439

Example 30

A study was performed to assess the amino acid profiles of the proteinsthat are involved in the high molecular weight lutein assembly from thesamples in Example 29. In order to compare the SEC peak area, responseto an independent measure of protein concentration, each of the sampleswere fractionated by SEC, and the individual fractions tested for aminoacids. In each case, 10×4-min fractions were collected (manually), overthe elution range of the hydrolysate (from about 20 min to about 60min), so that the volume of each fraction was 1.6 mL (i.e., 0.4 mL/min×4min). Each fraction was acid-digested (6 M HCl, 110° C., 22 h), and eachdigest was tested for eight amino acids (arginine [R], serine [S], ASX,[B]=aspartic acid [D]+asparagine [N], GLX [Z]=glutamic acid, [E]glutamine [Q], threonine [T], and glycine [G]) by reversed phase HPLC(RPLC) of their 9-fluorenylmethoxycarbonyl (FMOC) derivatives, using amodification of methods known in the art. The derivatised standard andsample solutions were tested for the FMOC derivatives of the six aminoacids by RPLC, using an Agilent model 1100 HPLC system (AgilentTechnologies, Wilmington, Del., USA) with an Agilent model G1321Afluorescence detector (FLD, Agilent Technologies, Wilmington, Del.,USA). The RPLC system was equipped with a 4.6×250 mm i.d., 5 μm, 120 Å,YMC-Pack ODS-AM reversed phase column (Waters, Milford, Mass., USA),maintained at 20° C. by a model G1316A thermostatted column compartment(Agilent Technologies, Wilmington, Del., USA). RPLC mobile phase A was65% (v/v) 0.05 M citric acid, pH 3.0 with NaOH, and 35% (v/v) ACN; RPLCmobile phase B was 20% (v/v) 0.05 M citric acid, pH 3.0 with NaOH, and80% (v/v) ACN. The flow rate was 0.5 mL/min, and the elution program wasas follows: 0.0-2.0 min, 0% B; 2.0-25.0 min, 0% B to 50% B (linear);25.1-48.0 min, 100% B; 48.1-63.0 min, 0% B. The FLD excitationwavelength was 262 nm, the emission wavelength was 310 nm, and the gainwas set at 10. The injection volume was 4 μL. The concentrations of R,S, B (D+N), Z (E+Q), T, and G in the sample solutions (i.e., preparedfrom the SEC fractions) were calculated by linear regression from therespective, individually-constructed, standard curves.

The table listed below displays the different amino acid ratios fordifferent proteins and the experimental groups. For example, NFDM,casein, whey, as well as the protein present in the aqueous fraction andnative protein. Of note, native protein refers to monomeric unaggregatedprotein, which has a molecular weight of approximately in the range of14-25 kD. Tube 1 refers to the sample with the highest amount of luteinassembly, while tube 6 refers to the sample having the lowest. They werechosen to optimally contrast the distribution of individual proteinswithin the assembly.

The data presented, suggest that a greater presence of casein is in thehigh molecular weight fraction than compared to native protein.

TABLE 15 Analysis of Proteins Associated with Assembly Amino acid ratio,w/w GLX/ASX ASX/Gly ASX/Ser ASX/Arg NFDM 2.76 3.59 1.39 2.10 Casein 3.203.90 1.23 1.90 Whey 1.58 5.96 2.09 4.42 β-casein 3.92 1.02 0.44 1.72Tube 1, high 2.47 1.22 0.53 2.59 MW protein Tube 1, native 1.77 1.411.05 5.51 protein Tube 6, high 2.71 0.97 0.42 1.96 MW protein Tube 6,native 1.76 1.48 1.04 5.32 protein

Example 31

A study was conducted to evaluate the presence of specific proteins thatassociate with the high molecular weight lutein complex. Commerciallyrepresentative nutritional powder batches (control and MDG incorporated)were reconstituted (152 g/L), and were analyzed before and aftercentrifugation (31,000×g/20° C./2 h). The components of these powdersare listed in the table of Example 22. Reversed phase HPLC was then usedto compare the distribution of individual proteins.

Determination of the proteins associated with lutein complex wasperformed on an Agilent model 1100 HPLC system (Agilent Technologies,Wilmington, Del., USA) with a model G1315A diode array detection (DAD)system (Agilent Technologies). The system was equipped with a 250 mm×4.6mm I.D., 5 μm, 120 Å, YMC-Pack ODS-AQ reversed-phase column (Waters,Milford, Mass., USA). The column was maintained at 40° C. duringanalyses by a model G1316A thermostatted column compartment(AgilentTechnologies). Direct determinations of intact proteins wereperformed on the same system equipped with a 250 mm×4.6 mm I.D., 5 μm,300 Å, Jupiter C18 reversed phase column (Phenomenex, Torrance, Calif.,USA), also with column temperature maintained at 40° C.

The reverse phase HPLC method used a binary gradient elution, withvacuum-degassed mobile phases A (water, Milli-Q Plus) and B (650 mL0.02M KH2PO4, pH 2.9, +175 mL acetonitrile+175 mL isopropanol). Theinjection volume was 2 μL, and the elution program was 0% B at 0.3mL/min from 0.0 to 20.0 min, 16% B at 0.5 mL/min from 20.2 to 40.0 min,100% B at 0.5 mL/min from 40.1 to 45.0 min, 0% B at 0.5 mL/min from 45.1to 59.0 min, and 0% B at 0.3 mL/min from 59.5 to 60.0 min. The intactprotein method also used a binary gradient elution, with vacuum-degassedmobile phases A (800 mL Milli-Q Plus water+200 mL acetonitrile+0.500 mLtrifluoroacetic acid) and B (250 mL Milli-Q Plus water+750 mLacetonitrile+0.500 mL trifluoroacetic acid). The elution program, with aconstant flow rate of 0.6 mL/min, was 0% B from 0.0 to 5.0 min 0 to 100%B from 5.0 to 40.0 min, 100% B from 40.0 to 45.0 min, and 0% B from 45.0to 60.0 min. The detection wavelengths were 214 nm and 280 nm, withreference at 590 nm. The injection volume was 10 μL, and the run timewas 60 min per injection.

The results demonstrated that the whole product had a significantdecreased presence of native α-casein in the MDG batches. This suggeststhat there may be enhanced calcium solubility within the MDGcomposition. Additionally, the aqueous fraction of MDG compositions hada significant increase in β-casein, which suggests that said protein maybe involved in the high molecular weight lutein assembly. The presenceof β-casein was further confirmed by the decrease in presence within thecream layer.

TABLE 16 Analysis of Proteins Associated with Assembly for CommerciallyRepresentative Nutritional Composition Sample Comparison ObservationInterpretation Whole Product Significantly Apparent increase indecreased presence of soluble calcium in A545 native alpha-casein in(alpha-caseins are A545 especially sensitive to soluble calcium) AqueousFraction Significantly Apparent increased (centrifugation increasedpresence of incorporation of supernatant) native beta-casein inbeta-casein into water A545 soluble, high MW assemblies, by MDG CreamLayer Slightly decreased Consistent with (extracted levels of nativeinterpretation of protein) beta-casein and Aqueous Fractionbeta-lactoglobulin in comparison A545

Example 32

A study was conducted to investigate the effects of variableconcentrations of MDG and HOSO, as well as the presence of differentproteins, on the formation of high molecular weight lutein assemblies.Model systems of different proteins: sodium caseinate, total milkprotein, whey protein concentrate and whey protein hydrolysate wereincorporated into the composition at 3.7 wt % of protein in the system.The details of the amounts of MDG, HOSO and lutein are detailed in thetable below. Samples were then centrifuged (31,000×g/20° C./1 h), andthe aqueous fractions were tested by SEC. The mobile phase used for SECwas 70% H₂O, 30% acetonitrile, and 0.1% trifluoracetic acid.

The data demonstrated that MDG compositions increased lutein assembly,and high molecular protein fractions across all groups compared to HOSOcompositions. The addition of whey protein hydolysate showed thegreatest amount of bound lutein. This suggests that it may beadvantageous to have whey protein hydrolysate within an MDG-basedcomposition. This finding is interesting in that it may be whey proteinhydrolysate's ability to aggregate that leads to enhanced luteinassembly. Furthermore, the SEC plots for sodium caseinate and total milkprotein suggest the presence of casein within the lutein complex.

TABLE 17 Model System Investigation III of Assembly Formation High MWlutein Aqueous assembly, Fraction Protein HOSO, MDG, Lutein, mAU-min/mg,protein >137 ingredient g/L g/L μg/L 476 nm kD, g/kg Sodium 6.67 0 6.9435.6 0.425 Caseinate 0 6.67 6.94 41.6 0.466 Total Milk 6.67 0 6.94 40.60.908 Protein 0 6.67 6.94 50.7 0.968 Whey 6.67 0 6.94 34.4 0.115 ProteinConcentrate 0 6.67 6.94 53.0 0.253 Whey 6.67 0 6.94 65.9 0.299 ProteinHydrolysate 0 6.67 6.94 97.0 0.453

Example 33

A study was conducted to evaluate the effect of relative centrifugalforce (RCF) on analyzing the high molecular weight lutein assembly inthe aqueous phase. The components of these powders are listed in thetable of Example 22. Samples were centrifuged, and the aqueous fractionswere tested by SEC. The mobile phase used for SEC was 70% H₂O, 30%acetonitrile, 0.1% trifluoracetic acid. The different RCFs used were1×g, 484×g, 1935×g, 7741×g and 31,000×g. These variables and others usedin the compositions are listed in the table below. The compositions werethen assessed for high molecular weight lutein assembly and the presenceof protein >137 kD as done above, as well as bound lipophile index.Bound lipophile index, a relative marker for of lipophilic nutrients inthe high molecular weight assembly, was determined as the peak ratio,280 nm/214 nm for protein >137 kD.

The results exhibit that the MDG compositions generated a high molecularweight lutein assembly, across all RCFs, which has significantly greaterwater solubility, and/or is more stable to external stress compared tocontrol compositions.

TABLE 18 Analysis of Variable Centrifugal Forces on the Assembly RCFHigh MW Bound Nutritional (20° C., lutein Protein >137 lipophile PowderBatch 2 hrs) assembly kD, g/kg index A544   1 × g 1839 5.23 0.299 A5452056 5.34 0.318 A544  484 × g 1969 5.27 0.306 A545 2274 5.60 0.327 A5441935 × g 1761 4.92 0.294 A545 2146 5.35 0.322 A544 7741 × g 729 2.740.232 A545 1163 3.50 0.277 A544 31,000 × g  244 1.07 0.184 A545 299 0.980.228

Example 34

A study was conducted to evaluate the effect of pH, during the SECprocess, on analyzing the presence of high molecular weight luteinassemblies. The components of these powders are listed in the table ofExample 22. In this experiment, neutral pH SEC was performed on MDG andcontrol samples. After the samples were reconstituted, they wereanalyzed via SEC under the conditions listed below. No centrifugationwas performed on samples prior to running on SEC.

The results demonstrate that under these conditions, the MDG-basedcompositions performed better than controls in the formation of the highmolecular weight lutein assembly, high molecular weight protein, andbound lipophile index. Additionally, this Example demonstrates that thedifference between MDG samples and control is not an artifact of theanalytical techniques used to assess the lutein assemblies.

TABLE 18 The Effect of pH on Assembly Characterization A544 (control)A545 (MDG) High MW lutein assembly, 372 419 mAU-min/mg injected High MWprotein, g/kg 3.61 3.73 Bound lipophile index 0.216 0.225

Example 35

A study was performed to investigate the effects of choline on theformation of high molecular weight lutein assemblies. Model systems of aprotein ingredient (whey protein concentrate or NFDM at 3.7 wt. % ofprotein), MDG (4.4 g/L) and lutein (6.7 mg/L). Additionally, differentexperimental groups also had either lecithin (949 mg/L), choline (45mg/L), or the combination thereof. Samples were then centrifuged(31,000×g/20° C./1 h), and the aqueous fractions were tested by SEC foranalysis on high molecular weight lutein assembly, high molecular weightprotein, and bound lipophile index. The data is presented in the tablebelow.

In summary, choline had a significant positive effect on the formationof high molecular weight lutein assemblies in the whey proteinconcentrate assemblies. It is hypothesized, without being bound to aparticular theory, that choline being a basic compound may interact withaspartic acid side chains, which are acidic. These interactions mayalter the 3-dimensional configuration of the major whey proteinβ-lactoglobulin such that the hydrophobic domains are more accessible tolutein binding.

It should also be noted that choline may have a selective enhancement oflutein incorporation, since the bound lipophile index actuallydecreased, even though lutein increased, when choline was present.

TABLE 19 Model System Investigation IV of Assembly Formation Bound HighMW lipophile lutein Aqueous index Protein assembly, Fraction (280ingredient mAU- protein > nm/214 (37 g protein Lecithin, Choline,min/mg, 137 kD, nm Tube per kg) mg/L mg/L 476 nm g/kg ratio) 1 WPC 0 031.2 0.105 0.157 2 NFDM 0 0 41.3 0.413 0.118 3 WPC 949 0 40.3 0.1430.163 4 NFDM 949 0 47.0 0.441 0.123 5 WPC 0 45 58.6 0.295 0.151 6 NFDM 045 50.4 0.443 0.131 7 WPC 949 45 61.7 0.338 0.146 8 NFDM 949 45 48.20.451 0.126

Example 36

A study was conducted to examine variations of commerciallyrepresentative nutritional composition batches. Samples were prepared asdetailed above, with composition with and without MDG. Samples werecentrifuged (7,7741×g/20° C./2 h), and the aqueous fractions were testedby SEC for analysis on high molecular weight lutein assembly, highmolecular weight protein, and bound lipophile index. The data ispresented in the table below.

In summary, the high molecular weight lutein assembly was 10-30% greaterin MGD compositions relative to control compositions. Additionally, theMDG compositions correlated with a high molecular weight protein peak.The estimated mass of the high molecular peak for these batches was inthe 240 kD range. There is also evidence that other lipophilic nutrients(e.g., Vitamin A, D, E, K, etc.) may be present within the highmolecular weight assembly.

TABLE 20 Analysis of Commercially Representative Nutritional Compositionin Assembly Formation Batch 017-1 017-2 017-3 017-4 017-5 Lutein/MDGControl PIF, w/ PIW PIF PIW, w/ addition lecithin choline Total lutein,2.18 1.99 2.06 1.80 1.88 mg/kg Lutein 2236 2464 2815 2844 2845 assembly,conc. Lutein 237 kD 240 kD 247 kD 237 kD 244 kD assembly, mass High MW5.93 6.34 6.75 6.92 6.87 protein, g/kg Bound 0.301 0.311 0.329 0.3270.329 lipophile index

Example 37

A study was conducted to investigate using a neutral pH as the SECrunning condition and its subsequent effect on the high molecular weightlutein assemblies during separation Samples were prepared as detailedabove, with composition with and without MDG. Samples were were testedby SEC for analysis on high molecular weight lutein assembly and boundlipophile index. SEC was performed using 0.05M HEPES, pH 7.0, and aSuperdex Peptide column. The data is presented in the table below.

The results indicate that the high molecular weight lutein assembly andprotein both increased in all MDG batches relative to control batches.This Example supports the integrity of the analytical techniques usedfor lutein assembly analysis.

TABLE 21 The Effect of pH on Assembly Characterization in CommerciallyRepresentative Nutritional Composition Batch 017-1 017-2 017-3 017-4017-5 Lutein/MDG Control PIF, w/ PIW PIF PIW, w/ addition lecithincholine Total lutein, 2.18 1.99 2.06 1.80 1.88 mg/kg Lutein 820 11701160 1190 1060 assembly, conc. Lipophile 7820 10,500 11,300 10,800 9510assembly, conc.

Example 38

A study was performed to evaluate the effects of choline on theformation of high molecular weight lutein assemblies by providingcholine at variable points during the composition. Mixtures of water,NFDM, MDG, lutein and choline chloride, wherein the choline was eitheradded to the MDG and lutein or to the NFDM slurry. Samples were thencentrifuged (31,000×g/20° C./1 h), ultrafiltrated through a 100 kfilter, and the aqueous fractions were tested by SEC for analysis onhigh molecular weight lutein assembly, high molecular weight protein,and native protein bound lutein. The supernatant was diluted 1:4 (v/v)prior to running on the SEC, with the SEC mobile phase (0.05M KH2PO4,0.15M NaCl, pH 7.5). The different parameters examined were highmolecular weight lutein assembly, protein >137 kD, and middle rangeprotein, which corresponds to the native protein, bound lutein.Additionally, ultrafiltrate analysis was performed on the luteinassembly and high molecular weight protein metrics. The data ispresented in the table below.

The enhancing effect of choline on lutein incorporation within highmolecular weight assemblies appears to be process dependent.Specifically, it was found that the bound lutein was significantlyhigher when choline was added to the NFDM slurry, as opposed to additionto the MDG and lutein.

TABLE 22 Model System Investigation V of Assembly Formation Cholineaddition point Added to Added to MDG + lutein NFDM slurry AqueousFraction, 214 nm 121 190 (protein metric) Aqueous Fraction, 476 nm 0.2650.330 (bound lutein metric) Aqueous Fraction, 476 nm, 0.0346 0.0455native protein range (native protein bound lutein metric) 100K UFpermeate, 214 nm 44.8 59.7 (protein metric) 100K UF permeate, 476 nm0.0343 0.0387 (lutein metric)

Example 39

A study was performed to evaluate the effects of variable components onthe formation of high molecular weight lutein assemblies. The differentcompositions include various proteins ingredients (NFDM,β-lactoglobulin, or bovine lactoferrin); varying amounts of MDG andHOSO, and lutein. The specific details of the aforementioned variablesare listed in the table below. Samples were then centrifuged(31,000×g/20° C./1 h), and tested for bound lutein via SEC.

The results demonstrate that both lutein assembly and protein >137 kDmetrics are increased with the presence of MDG within the composition.Interestingly, the combination of MDG and HOSO exhibited the greatestamount of lutein assembly formation and protein >137 kD, compared to MDGand HOSO alone. Additionally, the use of 3-lactoglobulin, or bovinelactoferrin, alone, as part of the composition showed the capability toform lutein assemblies and protein >137 kD.

TABLE 23 Model System Investigation VI of Assembly Formation Tube 1 2 34 5 6 7 8 Protein, 37 g NFDM NFDM NFDM NFDM NFDM NFDM βLG BLF protein/kgMDG, g/kg 6.64 6.62 3.31 1.66 0.826 0 6.62 6.62 HOSO, g/kg 0 0 3.31 4.965.79 6.62 0 0 Lutein, mg/kg 5.05 10.1 10.1 10.1 10.1 10.1 10.1 10.1MDG/lutein, 1315 655 328 164 81.8 0 655 655 w/w High MW 49.4 51.7 57.457.9 50.7 36.4 11.3 18.6 lutein assembly Protein > 137 0.413 0.407 0.4120.416 0.393 0.343 0.012 0.018 kD, g/kg Bound 0.136 0.137 0.147 0.1470.141 0.124 0.270 0.174 lipophile index

Example 40

A study was conducted to investigate the effects of variable componentson the formation of high molecular weight lutein assemblies. Thedifferent compositions include different protein ingredients (NFDM,β-lactoglobulin, or bovine lactoferrin) at 37 g of protein per kg, MDGat 6.62 g/kg, and lutein 10.1 mg/kg. The β-lactoglobulin, and bovinelactoferrin were tested both with and without centrifugation. Sampleswere then analyzed by SEC for high molecular weight lutein assembly,high molecular weight protein (>137 kD), either with centrifugation orwithout. If samples were centrifuged, they were done so at 31,000×g/20°C./1 h. Details of the different experimental group's ability to producehigh molecular weight lutein assemblies are shown in the table below.

The data revealed that both β-lactoglobulin and bovine lactoferrin werecapable of forming stable, high molecular weight lutein assemblies.Furthermore, these assemblies were still present after centrifugation,demonstrating the high solubility/stability of the bound luteinassemblies.

TABLE 24 Effect of Different Proteins on Assembly Formation in a ModelSystem Protein Sample High MW Bound (37 g prep lutein Protein >137lipophile Tube protein/kg) for SEC assembly kD, g/kg index 2 NFDMAqueous 51.7 0.407 0.137 7 β-LG Fraction 11.3 0.012 0.270 8 BLF 18.60.018 0.174 7 β-LG Whole 66.4 0.144 0.324 8 BLF suspension 60.5 0.1320.305

Example 41

A study was conducted to examine the effects of variable components onthe formation of high molecular weight lutein assemblies. Samples weresubjected to appropriate thermal processing during which the compositionis pre-heated to 165-185° F. and then passed through homogenizers, firstat 1000 psig and then at 4000 psig with NFDM at 37 g of protein/L, withlutein at 10.1 mg/L, and with varying amounts of HOSO and/or MDG. Afterhomogenization the each variable is subjected to a hightemperature-short time (HTST) process of about 165-185° F. and a 16second hold and then cooled to about 40° F. All samples were then testedby SEC, either following centrifugation (31,000×g/20° C./1 h) or withoutcentrifugation. The data is listed in the table below.

The results demonstrate that the combination of MDG and HOSO led to thehighest amount of lutein assembly formation, as well as protein >137 kDprior to centrifugation.

TABLE 25 Model System Investigation VII of Assembly Formation High MWBound HOSO, MDG, lutein Protein >137 lipophile Tube g/L g/L assembly kD,g/kg index 1 6.62 0 216 0.706 0.258 2 2.21 0 102 0.464 0.195 3 0 6.62167 0.667 0.226 4 0 2.21 65 0.384 0.165 5 3.31 3.31 195 0.712 0.246 64.97 1.65 208 0.747 0.247 results of testing without centrifugation 16.62 0 612 2.06 0.258 2 2.21 0 242 1.18 0.198 3 0 6.62 648 2.27 0.246 40 2.21 260 1.66 0.165 5 3.31 3.31 970 2.96 0.286 6 4.97 1.65 888 2.570.297

Example 42

A study was conducted to examine the effects of variable components onthe formation of high molecular weight lutein assemblies using acommercially available nutritional powder. Samples were tested (withoutdilution or centrifugation) for bound lutein and related attributes bySEC. The attributes are compared below, and the bound lutein values(peak area at 476 nm, per mg injected) are also compared in the tablebelow.

The results demonstrate that the combination of MDG provided the highestamount of the metrics listed below relative to controls. Specifically,MDG/choline outperformed all other groups across all metrics.

TABLE 26 Effect of Different Variables on Assembly Formation in aCommercially Representative Nutritional Composition Lutein, High MWBound mg/kg lutein Protein >137 lipophile Variation (as fed) assemblykD, g/kg index Control 2.47 6995 13.8 0.373 MDG/PIF 2.47 9189 16.3 0.419MDG/PIW 2.35 10,566 17.8 0.441 MDG/lecithin 2.44 10,298 17.2 0.446MDG/choline 2.11 10,937 18.1 0.448

Example 43

A study was conducted to examine the effects of variable components onthe formation of high molecular weight lutein assemblies. Each variablewas subjected to appropriate thermal processing during which thecomposition is pre-heated to 165-185° F. and then passed throughhomogenizers, first at 1000 psig and then at 4000 psig. Afterhomogenization, each variable was subjected to a high temperature-shorttime (HTST) process of about 165-185° F. and a 16 second hold and thencooled to about 40° F. Samples were homogenized and pasteurized with thefollowing parameters of protein (“P”=nonfat dry milk, at 10% w/w),lutein (“L”), HOSO (“H”), MDG (“M”), lecithin (“Le”), and/or choline(“C”). These samples were tested for bound lutein and related attributesby size exclusion chromatography (SEC). Each variable was tested bothbefore and after centrifugation (at 31,000×g/20 C/1 h).

The results demonstrate that the presence of MDG led to the formation oflutein assembly formation and protein >137 kD.

TABLE 27 Model System Investigation VIII of Assembly Formation High MWBound lutein Protein >137 lipophile Variable Sample assembly kD, g/kgindex PLH Whole 779 2.21 0.293 PLH Supernatant 328 (42%) 0.97 (44%)0.281 PLM Whole 439 1.28 0.273 PLM Supernatant 321 (73%) 0.90 (75%)0.286 PLMLe Whole 845 3.03 0.253 PLMLe Supernatant 239 (28%) 1.13 (37%)0.215 PLMC Whole 432 1.27 0.269 PLMC Supernatant 323 (75%) 0.98 (77%)0.276

Example 44

A study was conducted to examine the effects of variable components onthe formation of high molecular weight lutein assemblies. Samples weresubjected to appropriate thermal processing during which the compositionwas pre-heated to 165-185° F. and then passed through homogenizers,first at 1000 psig and then at 4000 psig with NFDM at 37 g of protein/L,with lutein at 10.1 mg/L, and with varying amounts of HOSO and/or MDG.After homogenization each variable was subjected to a hightemperature-short time (HTST) process of about 165-185° F. and a 16second hold, and then cooled to about 40° F. All samples were thentested by SEC, either following centrifugation (31,000×g/20° C./1 h) orwithout centrifugation. The data is listed in the table below.

Samples prepared with lutein, a lipid (HOSO or MDG), and varying levelsof protein (nonfat dry milk), were tested for bound lutein and relatedattributes by size exclusion chromatography (SEC). Specifically, theparameters investigated were high molecular weight lutein assembly,protein >137 kD, and bound lipophile index. The results for samplesprepared with and without centrifugation (31,000×g; 20 C; 1 h) arecompared below.

The results demonstrate that both of MDG and HOSO led to the higheramounts of lutein assembly formation, protein >137 kD, and boundlipophilic index with increasing amounts of protein. Additionally, thepresences of MDG resulted mostly in higher amounts of the aforementionedmetrics, relative to HOSO. Interestingly, MDG outperformed HOSO moresignificantly as the protein was increased via all metrics.

TABLE 28 Model System Investigation VIII of Assembly Formation ProteinHigh MW Bound (NFDM), Lipid, lutein Protein >137 lipophile Tube g/kg 4.4g/kg assembly kD, g/kg index 1 24.1 HOSO 107 2.17 0.103 2 24.1 MDG 1222.11 0.107 3 48.2 HOSO 218 4.15 0.110 4 48.2 MDG 232 4.15 0.112 5 96.4HOSO 506 10.1 0.121 6 96.4 MDG 616 11.6 0.125 Tested aftercentrifugation at 31,000 × g and at 20° C. for 1 hr 1 24.1 HOSO 49.80.884 0.096 2 24.1 MDG 53.8 1.02 0.098 3 48.2 HOSO 108 2.44 0.102 4 48.2MDG 118 2.48 0.101 5 96.4 HOSO 287 5.31 0.119 6 96.4 MDG 309 5.47 0.125

Example 45

A study was conducted to examine the presence of tocopherol within thehigh molecular weight assembly. The components of the tested powders arelisted in the table of Example 22. All samples were formulated and thenthen tested by SEC, following centrifugation (31,000×g/20° C./3 h). Thedata is listed in the table below.

TABLE 29 Analysis of Tocopherol within the Assembly NutritionalTocopherols (α-, γ-, and δ-) in Powder Aqueous Fraction, approximateA544 (control) 1.51 mg/kg A545 (MDG) 1.69 mg/kg

The results demonstrated that the aqueous fractions of commerciallyavailable nutritional powders with MDG contained tocopherols are at alevel that is 11% greater than control compositions. The aqueousfraction concentrations (˜1.69 and ˜1.51 mg/kg, respectively) appear torepresent an appreciable proportion of the total tocopherol within thecommercially available nutritional composition (approximately 20-30%).The tocopherol solubility provided by the MDG within the composition isin stark contrast to the reported solubility limit of vitamin E in water(20.9 mg/kg at 33° C.). This suggests that the assemblies comprising MDGenhance the aqueous solubility of tocopherol.

Example 46

TABLE 30 Analysis of Casein Associated with the Assembly Currentcommercial Protein system composition Composition 64% nonfat dry milkNFDM from Holstein- NFDM from Guernsey (NFDM), 36% WPC Friesian breedsbreed Total protein 2.07 g per 100 kcal 2.07 g per 100 kcalconcentration Total β-casein 0.36 g per 100 kcal 0.36 g per 100 kcalconcentration Major genetic variant A1 A2 of β-casein

Table 31 below shows partial composition information (protein content)for a commercial liquid nutritional composition designed for individualswith diabetes, compared with an exemplary liquid nutritional compositionaccording to the embodiments disclosed herein. Both illustrate liquidnutritional product with varying sources of caseinates, and have acaseinate, SPC combination. The beta-casein utilized in the embodimenton the right is sourced from Bos indicus has A2 as the major geneticvariant.

TABLE 31 Partial Composition Information of Commercially AvailableNutritional Composition Current commercial Protein system compositionComposition 80% casemates, Caseinates from Caseinates from 20% SPC Bostaurus milk, Bos indicus milk Holstein-Friesian breeds Total protein 76g per liter 76 g per liter concentration Total β-casein 22 g per liter22 g per liter concentration Major genetic variant A1 A2 of β-casein

A nutritional composition intended for infants and suitable forutilizing the protein disclosed herein is described in Table 32 below.The 14 grams of protein per liter of nutritional composition comprisesabout 18% by weight bovine beta-casein.

TABLE 32 Nutritional Composition Ingredients for Example 46 and otherExemplified Compositions Amount per Ingredient Name 1000 Kg batchKg/g/mg Ingredient Water Q.S. Kg Nonfat Milk 83.00 Kg Lactose 51.82 KgHigh Oleic Safflower Oil 13.06 Kg Soy Oil 10.50 Kg Coconut Oil 9.256 KgGalacto-oligosaccharides 8.630 Kg Whey Protein Concentrate 6.075 Kg 5%Potassium Hydroxide 2.494 Kg KOH 124.7 g Potassium Citrate 536.6 gCalcium Carbonate 449.9 g Ascorbic Acid 446.9 g ARASCO MortierellaAlpina Oil 359.3 g Soy Lecithin 339.7 g Myverol 18-06 339.7 gNucleotide-Choline Premix 293.2 g Choline Bitartrate 51.75 g Cytidine5′-Monoohosohate 30.49 g Disodium Guanosine 5′-Monophosphate 15.64 gDisodium Uridine 5′-Monophosphate 13.15 g Adenosine 5′-Monophosphate11.60 g Potassium Chloride 199.9 g Magnesium Chloride 154.0 gVit/Min/Taur Premix 149.9 g Taurine 45.83 g m-lnositol 33.28 g ZincSulfate 15.35 g Niacinamide 9.781 g Calcium Pantothenate 5.865 g FerrousSulfate 5.131 g Cupric Sulfate 1.800 g Thiamine Chloride HCI 1.518 gRiboflavin 669.3 g Pvridoxine HCI 613.1 mg Folic Acid 206.1 mg ManganeseSulfate 174.6 mg Biotin 59.21 mg Sodium Selenate 35.51 mg Cyanocobalamin4.722 mg DHASCO Crypthecodinium Cohnii Oil 131.0 g Choline Chloride123.7 g Seakem GP-359 120.0 g Ultra-Micronized Tricalcium Phosphate103.2 g Potassium Phosphate Monobasic 90.6 g Vitamin A, D3, E, K1 Premix64.7 g RRR Aloha-Tocophervl Acetate 9.063 g Vitamin A Palmitate 1.725 gVitamin K1 (Phylloquinone) 100.3 mg Vitamin D3 14.06 mg Ferrous Sulfate60.9 g Seakem RLC Carrageenan 60.0 g Carotenoid Premix 57.1 g Lutein119.9 mg Lycopene 119.9 mg Beta- Carotene 25.98 mg Sodium Chloride 40.1g Citric Acid (Processing Aid) 29.8 g L-Carnitine 3.62 g Riboflavin 2.18g

A nutritional composition suitable for utilizing the protein disclosedherein is described in Table 33 below. The 34 grams of protein per literof nutritional composition comprises about 25% by weight bovinebeta-casein.

TABLE 33 Nutritional Composition Ingredients for Example 46 and otherExemplified Compositions Ingredient Kg per 1000 Kg Water Q.S. Sucrose75.0 Milk Protein Isolate (Alapro 4900- Fonterra) 34.9 Soy Oil 8.0 SoyProtein Isolate 3.7 Potassium Citrate 3.3 Phytosterol Ester 3.2 AvicelCL611 3.0 Magnesium Phosphate Dibasic 1.92 N&A Vanilla Flavor 1.80 m-TCP1.75 N&A Dairv Cream Flavor 32122 1.30 Soy Lecithin 1.0 MagnesiumChloride 1.15 Sodium Chloride 0.75 Choline Chloride 0.53 Ascorbic Acid0.377 Potassium Chloride 0.309 Potassium Hydroxide 45% (Processing Aid)0.262 WSV Premix 0.185 Corn Starch (Processing Aid) 0.0631143 Dextrose(Processing Aid) 0.062977 Niacinamide 0.0274725 d-Calcium Pantothenate0.0177239 Thiamine Chloride Hydrochloride 0.0045325 PyridoxineHydrochloride 0.0043625 Riboflavin 0.0035335 Folic Acid 0.0006739 Biotin0.0005467 Cyanocobalamin 0.00006318 Carrageenan (Viscarin SA-359) 0.180UTM/TM Premix 0.150 Zinc Sulfate, Monohydrate 0.0837855 Maltodextrin(Processing aid) 0.0346267 Manganese Sulfate, Monohydrate 0.0207663Citric acid, Anhydrous 0.009258 Chromium Chloride, Hexahydrate 0.0007288Sodium Molybdate, Dihydrate 0.0005882 Sodium Selenate, Anhydrous0.0002465 Vitamin ADEK Premix 0.135 Coconut Oil (Processing Aid)0.0799578 Vitamin E (dl-Alpha Tocophervl Acetate) 0.0495827 Vitamin APalmitate 0.0051266 Phylloquinone 0.000297 Vitamin D3 0.00003594Sucralose Liquid (25%) 0.10 Seakem CM514 0.090 Acesulfame potassium0.050 Potassium Iodide 0.00023

In certain exemplary embodiments disclosed herein, the nutritionalcompositions are administered (or consumed) orally as needed to providethe desired level of nutrition. In certain of these embodiments, thenutritional compositions are administered (or consumed) in the form ofone to two servings daily or in one or two or more divided doses daily.In certain embodiments, when the nutritional composition is a liquid,the serving may be 150 milliliters to 500 milliliters. In certain otherembodiments, when the nutritional composition is a liquid, the servingis 237 milliliters (˜8 fl. oz.). In other embodiments, when thenutritional composition is a liquid, the serving is 177 milliliters to414 milliliters (˜6 fl. oz. to ˜14 fl. oz.). In yet other embodiments,when the nutritional composition is a liquid, the serving is 207milliliters to 266 milliliters (˜7 fl. oz. to ˜9 fl. oz.). Variouscalorie contents may be associated with each serving of the nutritionalcompositions according to the first, second and third embodimentsdisclosed herein, typically from 25 to 500 Kcal, including 50 to 400Kcal; 100 to 350 Kcal or 150 to 350 Kcal per serving. Alternatively, aserving may be construed as any amount which is intended to be consumedin one sitting or within one hour or less.

To the extent that the term “includes” or “including” is used in thespecification or the claims, it is intended to be inclusive in a mannersimilar to the term “comprising” as that term is interpreted whenemployed as a transitional word in a claim. Furthermore, to the extentthat the term “or” is employed (e.g., A or B) it is intended to mean “Aor B or both.” When the applicants intend to indicate “only A or B butnot both” then the term “only A or B but not both” will be employed.Thus, use of the term “or” herein is the inclusive, and not theexclusive use. See Bryan A. Garner, A Dictionary of Modern Legal Usage624 (2d. Ed. 1995). Also, to the extent that the terms “in” or “into”are used in the specification or the claims, it is intended toadditionally mean “on” or “onto.” Furthermore, to the extent the term“connect” is used in the specification or claims, it is intended to meannot only “directly connected to,” but also “indirectly connected to”such as connected through another component or components.

While the present application has been illustrated by the description ofembodiments thereof, and while the embodiments have been described inconsiderable detail, it is not the intention of the applicants torestrict or in any way limit the scope of the appended claims to suchdetail. Additional advantages and modifications will readily appear tothose skilled in the art. Therefore, the application, in its broaderaspects, is not limited to the specific details, the representativeapparatus, and illustrative examples shown and described. Accordingly,departures may be made from such details without departing from thespirit or scope of the applicant's general inventive concept.

Examples 47-53

Examples 47-49 illustrate soy-based powdered infant formulas of thepresent disclosure, the ingredients of which are listed in Table 34. Allingredient amounts are listed as kg per 1000 kg batch, unless otherwisespecified.

TABLE 34 Nutritional Composition Ingredients for Examples 47-49 andother Exemplified Compositions Ingredient Example 1 Example 2 Example 3Corn Syrup 504.1 504.1 504.1 Soy Protein Isolate (5% DH) 144.8 144.8144.8 Sunflower Oil 112.5 112.5 112.5 Sucrose 98.3 98.3 98.3 Soy Oil83.9 83.9 83.9 Coconut Oil 75.6 75.6 75.6 Fructooligosaccharides 17 1717 Potassium Citrate 16.5 16.5 16.5 Calcium Phosphate 16.4 16.4 16.4Sodium Chloride 3.8 3.8 3.8 Arachidonic Acid Oil 3 3 3 MagnesiumChloride 2.8 2.8 2.8 L-Methionine 1.7 1.7 1.7 Ascorbic Acid 1.1 1.1 1.1Docosahexaenoic Acid Oil 1.1 1.1 1.1 MDG Oil 621.0 g 590.3 g 558.9 gLutein 945.0 mg 346 mg 1.1 g Choline Chloride 507.7 g 507.7 g 507.7 gTaurine 457.5 g 457.5 g 457.5 g Inositol 353.0 g 353.0 g 353.0 gAscorbyl Palmitate 347.5 g 347.5 g 347.5 g Ferrous Sulfate 319.2 g 319.2g 319.2 g Mixed Tocopherols 157.2 g 157.2 g 157.2 g L-Carnitine 112.7 g112.7 g 112.7 g Niacinamide 97.9 g 97.9 g 97.9 g D-Alpha-TocopherylAcetate 78.8 g 78.8 g 78.8 g Calcium D-Pantothenate 58.7 g 58.7 g 58.7 gZinc 56.0 g 56.0 g 56.0 g Iron 16.9 g 16.9 g 16.9 g Thiamine 15.2 g 15.2g 15.2 g Vitamin A Palmitate 14.8 g 14.8 g 14.8 g Copper 7.2 g 7.2 g 7.2g Riboflavin 6.7 g 6.7 g 6.7 g Pyridoxine Hydrochloride 6.1 g 6.1 g 6.1g Folic Acid 2.1 g 2.1 g 2.1 g Potassium Iodide 1.1 g 1.1 g 1.1 gPhylloquinone 857.1 mg 857.1 mg 857.1 mg Vitamin D3 47 mg 103.7 mg 324mg Lycopene 980.0 mg 980.0 mg 980.0 mg Biotin 592.5 mg 592.5 mg 592.5 mgBeta-Carotene 215.6 mg 215.6 mg 215.6 mg Selenium 147.0 mg 147.0 mg147.0 mg Cyanocobalamin 71.3 mg 71.3 mg 71.3 mg

Examples 50-52 illustrate milk-based powdered infant formulas of thepresent disclosure, the ingredients of which are listed in Table 35. Allingredient amounts are listed as kg per 1000 kg batch, unless otherwisespecified.

TABLE 35 Nutritional Composition Ingredients for Examples 50-52 andother Exemplified Compositions Ingredient Example 4 Example 5 Example 6Skim Milk 2057.3 2057.3 2057.3 Lactose 414.1 414.1 414.1 Sunflower Oil111.1 111.1 111.1 Soy Oil 83.3 83.3 83.3 Coconut Oil 74.6 74.6 74.6Galactooligosaccharides 72.3 72.3 72.3 Whey Protein Concentrate 49.449.4 49.4 Whey Protein Hydrolysate 5.6 5.6 5.6 (<25% DH) PotassiumCitrate 3.4 3.4 3.4 Arachidonic Acid Oil 2.9 2.9 2.9 Calcium Carbonate2.6 2.6 2.6 Potassium Hydroxide 2.3 2.3 2.3 Soy Lecithin 1.7 1.7 1.7Docosahexaenoic Acid Oil 1.6 1.6 1.6 Ascorbic Acid 1.5 1.5 1.5 CholineBitartrate 1.2 1.2 1.2 Magnesium Chloride 1.2 1.2 1.2 MDG Oil 1.1 1.0440.99 Lutein 2.65 g 1.5 g 364 mg Sodium Chloride 1 1 1 Taurine 428 g 428g 428 g L-Tryptophan 277 g 277 g 277 g Cytidine 5′-Monophosphate 257 g257 g 257 g Myo-Inositol 257 g 257 g 257 g Ascorbyl Palmitate 200 g 200g 200 g Disodium Guanosine 187 g 187 g 187 g 5′-Monophosphate DisodiumUridine 163 g 163 g 163 g 5′-Monophosphate Ferrous Sulfate Monohydrate130.6 g 130.6 g 130.6 g RRR-Alpha-Tocopheryl Acetate 119.7 g 119.7 g119.7 g Tocopherol-2 106.7 g 106.7 g 106.7 g Zinc Sulfate Monohydrate104 g 104 g 104 g Adenosine 5′-Monophosphate 98 g 98 g 98 g FerrousSulfate 88 g 88 g 88 g Mixed Tocopherols 77 g 77 g 77 g Niacinamide 59 g59 g 59 g L-Carnitine 43 g 43 g 43 g Calcium D-Pantothenate 42 g 42 g 42g Retinyl Palmitate 13 g 13 g 13 g Copper Sulfate 11 g 11 g 11 g ThiaminHydrochloride 9 g 9 g 9 g Pyridoxine Hydrochloride 5 g 5 g 5 gRiboflavin 5 g 5 g 5 g Manganese Sulfate Monohydrate 3 g 3 g 3 g FolicAcid 796 mg 796 mg 796 mg Beta-Carotene 770 mg 770 mg 770 mgPhylloquinone 618 mg 618 mg 618 mg Potassium Iodide 550 mg 550 mg 550 mgD-Biotin 238 mg 238 mg 238 mg Sodium Selenate 117 mg 117 mg 117 mgVitamin D3 50 mg 93 mg 295 mg Cyanocobalamin 8 mg 8 mg 8 mg

Example 53 illustrates a ready-to-feed milk-based liquid infant formulaof the present disclosure, the ingredients of which are listed in Table36. All ingredient amounts are listed as kg per 1000 kg batch, unlessotherwise specified.

TABLE 36 Nutritional Composition Ingredients for Example 53 and otherExemplified Compositions Ingredient Example 7 Water Q.S. Lactose 48.2Skim Milk 22.1 High Oleic Safflower Oil 13.3 Soy Oil 10.2 Coconut Oil9.7 Galactooligosaccharides 9.4 Whey Protein Concentrate 6.3 PotassiumHydroxide 166 g Calcium Carbonate 747 g Potassium Citrate 650 g AscorbicAcid 610 g Arachidonic Acid Oil 410 g Distilled Monoglycerides 347 gLecithin 350 g Cytidine 5′-Monophosphate 32.8 g Disodium Guanosine5′-Monophosphate 16.8 g Disodium Uridine 5′-Monophosphate 14.1 gAdenosine 5′-Monophosphate 12.5 g Docosahexaenoic Acid Oil 225 g MDG Oil210 g Potassium Phosphate 185 g Taurine 57 g Myo-Inositol 48 gL-Tryptophan 19 g Niacinamide 6 g Calcium D-Pantothenate 5 g ThiamineHydrochloride 1.4 g Riboflavin 1.3 g Pyridoxine Hydrochloride 750 mgFolic Acid 145 mg D-Biotin 36 mg Cyanocobalamin 5.4 mg Carrageenan 175 gSodium Citrate 174 g Sodium Chloride 145 g Magnesium Chloride 126 gFerrous Sulfate 66.5 g Choline Chloride 45 g Vitamin E Acetate 21 gVitamin A Palmitate 2 g Phylloquinone 131.6 mg Vitamin D3 16 mgMagnesium Sulfate 32 g Zinc Sulfate 17.9 g L-Carnitine 5.5 g CupricSulfate 2 g Lutein 160 mg Beta-Carotene 80 mg Manganese Sulfate 110 mg

Example 54

In this Example, the effect of using an activated MDG oil component oran MDG-protected premix on the absorbance of triglycerides and fattyacids in a nutritional composition is evaluated and compared to theabsorbance of triglycerides and fatty acids in a nutritional compositionwithout MDG oil. The fatty acids that were measured for this testincluded palmitic acid, linoleic acid, and alpha-linolenic acid.

The following nutritional compositions were tested:

Triglyceride oil control (AET-C): high oleic safflower oil was added tothe protein-in-fat slurry prior to addition to the remainder of thecomposition;

MDG-protected premix (AET-1): MDG oil was premixed with lecithin for 5minutes; this premix was added to the protein-in-fat slurry to form anMDG-protected premix prior to addition to the remainder of thecomposition;

Activated MDG oil component (AET-2): MDG oil was added to theprotein-in-water slurry to form an activated MDG oil component prior toaddition to the remainder of the composition;

MDG oil, not activated (AET-3): MDG oil was added to the protein-in-fatslurry prior to addition to the remainder of the composition; and

Activated MDG oil component with choline (AET-4): MDG oil was premixedwith choline for 5 minutes; this premix was added to theprotein-in-water slurry to form an activated MDG oil component prior toaddition to the remainder of the composition.

TABLE 37 Nutritional Composition Ingredients for Example 54 and otherExemplified Compositions Sample ID: AET-C AET-1 AET-2 AET-3 AET-4Amount/ Amount/ Amount/ Amount/ Amount/ Ingredient Name: 1000 kg Unit1000 kg Unit 1000 kg Unit 1000 kg Unit 1000 kg Unit Water Q.S. kg Q.S.Kg Q.S. kg Q.S. kg Q.S. kg Lactose 49.27 kg 49.27 Kg 49.27 kg 49.27 kg49.27 kg Non-Fat Dry Milk 22.06 kg 22.06 Kg 22.06 kg 22.06 kg 22.06 kgHigh Oleic 13.44 kg 12.78 Kg 12.78 kg 12.78 kg 12.78 kg Safflower OilSoy Oil 10.06 kg 9.57 Kg 9.57 kg 9.57 kg 9.57 kg Coconut Oil 9.60 kg9.13 Kg 9.13 kg 9.13 kg 9.13 kg Galactooligo- 9.40 kg 9.40 Kg 9.40 kg9.40 kg 9.40 kg saccharides Whey Protein 6.31 kg 6.31 Kg 6.31 kg 6.31 kg6.31 kg Concentrate MDG Oil 0 kg 1.62 Kg 1.62 kg 1.62 kg 1.62 kgAscorbic Acid 611.99 g 611.99 G 611.99 g 611.99 g 611.99 g CalciumCarbonate 559.70 g 559.70 G 559.70 g 559.70 g 559.70 g Potassium Citrate545.40 g 545.40 G 545.40 g 545.40 g 545.40 g ARASCO 395.30 g 395.30 G395.30 g 395.30 g 395.30 g Mortierella Alpina Oil Potassium Hydroxide,45% 368.00 g 368.00 G 368.00 g 368.00 g 368.00 g Lecithin 346.99 g346.99 G 346.99 g 346.99 g 346.99 g Myverol 346.99 g 346.99 G 346.99 g346.99 g 346.99 g Nucleotide- 314.15 g 314.15 G 314.15 g 314.15 g 314.15g Choline Premix DHASCO 208.80 g 208.80 G 208.80 g 208.80 g 208.80 gCrypth. Cohnii Oil Vitamin/Mineral/ 176.00 g 176.00 G 176.00 g 176.00 g176.00 g Taurine Premix Carrageenan 175.26 g 175.26 G 175.26 g 175.26 g175.26 g Sodium Citrate 163.00 g 163.00 G 163.00 g 163.00 g 163.00 gSodium Chloride 145.30 g 145.30 G 145.30 g 145.30 g 145.30 g MagnesiumChloride 126.50 g 126.50 G 126.50 g 126.50 g 126.50 g PotassiumPhosphate 117.60 g 117.60 G 117.60 g 117.60 g 117.60 g Dibasic Ferroussulfate 66.50 g 66.50 G 66.50 g 66.50 g 66.50 g Choline Chloride 44.97 g44.97 G 44.97 g 44.97 g 44.97 g Vitamin A, D3, E, K1 35.80 g 35.80 G35.80 g 35.80 g 35.80 g Magnesium Sulfate 27.40 g 27.40 G 27.40 g 27.40g 27.40 g Zinc Sulfate 17.90 g 17.90 G 17.90 g 17.90 g 17.90 g Lutein12.35 g 12.35 G 12.35 g 12.35 g 12.35 g L-Carnitine 5.50 g 5.50 G 5.50 g5.50 g 5.50 g Cupric Sulfate 1.90 g 1.90 G 1.90 g 1.90 g 1.90 gManganese Sulfate 110.00 mg 110.00 Mg 110.00 mg 110.00 mg 110.00 mgCitric Acid As As As As As needed needed needed needed needed PotassiumIodide As As As As As needed needed needed needed needed Sodium SelenateAs As As As As needed needed needed needed needed Potassium Chloride AsAs As As As needed needed needed needed needed

Male Sprague Dawley rats weighing between 280 and 330 grams were all feda normal commercially available Purina Rat Chow for one week. Rats werethen fasted overnight prior to surgery, and under anesthesia, alaparotomy was performed, and the intestinal lymph duct was cannulatedaccording to the procedure of Tso et al., “The Absorption of Lipid andLipoprotein Synthesis,” Lipid Research Methodology, Chapter 5: 191-216(1984) Alan R. Liss, Inc., NY, N.Y., hereby incorporated by reference tothe extent consistent herewith. The superior mesenteric artery wasisolated, but not occluded. A silicon infusion tube (1.6 mm OD) wasplaced in the stomach for future infusion of a test sample. The fundicincision was closed by a purse string suture. The rats were allowed torecover for 24 hours before infusion began.

The rats were randomly assigned to receive one of the five nutritionalcompositions being tested. (The number of rats in each group, e.g.“(n=7),” is shown in the key for FIG. 18.) The animals wereintragastrically infused 24 hours after surgery with 3 ml of theirrespective test nutritional compositions.

Lymph was collected in pre-cooled tubes for 1 hour to measure fastinglevels of triglycerides, palmitic acid, linoleic acid, and alphalinolenic acid before the nutritional composition infusions began. Lymphwas then collected in pre-cooled tubes hourly for 6 hours afterinitiation of infusion. At the end of the experiment, the rats weresacrificed by exsanguination.

The lymph lipids were extracted and analyzed for triglycerides, palmiticacid, linoleic acid, and alpha-linolenic acid using high-performanceliquid chromatography (Craft Technologies, Wilson, N.C.).

FIG. 18 shows the lymphatic output of triglycerides over a period of 6hours following infusion of the liquid nutritional compositions. FIG. 19shows the lymphatic absorption of palmitic acid over a 6 hour periodfollowing infusion of the liquid nutritional compositions. FIG. 20 showsthe lymphatic absorption of linoleic acid over a 6 hour period followinginfusion of the liquid nutritional compositions. FIG. 21 shows thelymphatic absorption of alpha-linolenic acid over a period of 6 hoursfollowing infusion of the liquid nutritional compositions.

As shown in FIG. 18, the mean fasting lymphatic triglyceride output forall groups of rats varied between about 2.0 and 3.5 mg/hr. In allgroups, lymphatic triglyceride output increased after infusion of eachnutritional composition began and reached a maximum output of about 8 toabout 17.5 mg/hr two hours after infusion began. The control group(AET-C) and the group receiving MDG oil that was not activated (AET-3)showed modest increases in the lymphatic triglyceride output, with atriglyceride output of about 8 to about 9 mg/hr two hours after infusionbegan. However, the groups receiving either an activated MDG component(AET-2 and AET-4) or an MDG-protected premix (AET-1) showed greaterlymphatic triglyceride output, with a triglyceride output of about 13 toabout 17.5 mg/hr two hours after infusion of the nutritionalcompositions began.

As shown in FIG. 18, the mean fasting lymphatic triglyceride output forall groups of rats varied between about 2.0 and 3.5 mg/hr. In allgroups, lymphatic triglyceride output increased after infusion of eachnutritional composition began and reached a maximum output of about 8 toabout 17.5 mg/hr two hours after infusion began. The control group(AET-C) and the group receiving MDG oil that was not activated (AET-3)showed modest increases in the lymphatic triglyceride output, with atriglyceride output of about 8 to about 9 mg/hr two hours after infusionbegan. However, the groups receiving either an activated MDG component(AET-2 and AET-4) or an MDG-protected premix (AET-1) showed greaterlymphatic triglyceride output, with a triglyceride output of about 13 toabout 17.5 mg/hr two hours after infusion of the nutritionalcompositions began.

FIG. 20 shows the lymphatic absorption of linoleic acid for 6 hoursfollowing infusion of the five nutritional compositions. The lymphaticabsorption of linoleic acid increased in all groups during the first twohours and fell to a steady state from about 4 to 6 hours after infusionof the nutritional compositions began. However, there was a markedincrease in the lymphatic absorption of linoleic acid two hours afterinfusion began for the nutritional compositions containing some form ofMDG oil (AET-1 to AET-4), when compared with the nutritional compositioncontaining the triglyceride control with no MDG oil (AET-C).

FIG. 21 shows the lymphatic absorption of alpha-linolenic acid for 6hours following infusion of the five nutritional compositions. Thelymphatic absorption of alpha-linolenic acid increased in all groupsduring the first two hours and fell to a steady state from about 4 to 6hours after infusion of the nutritional compositions began. However,there was a marked increase in the lymphatic absorption ofalpha-linolenic acid two hours after infusion began for the nutritionalcompositions containing some form of MDG oil (AET-1 to AET-4), whencompared with the nutritional composition containing the triglyceridecontrol with no MDG oil (AET-C).

Example 55

In this Example, the nutritional compositions evaluated in Example 1were further tested for lutein that is not solubilized by the fat in thenutritional compositions. The five compositions, AET-C, AET-1, AET-2,AET-3 and AET-4, were each subjected to high speed centrifugation(31,000×g; 20° C.; 4 h). The aqueous fraction of each centrifugednutritional composition was diluted 2:8 with water, and the dilutedaqueous fractions were filtered through 0.45 μm PTFE membranes. Thevisible absorbance of the diluted aqueous fractions was measured at 476nm (a lutein absorbance maximum). The absorbance readings(milliabsorbance units per gram of aqueous fraction (mAU/g)) arereported in Table 3 below, along with a ranking of the in vivo luteinresponse for the same compositions tested in Example 1.

TABLE 38 Relationship between Lutein in the Aqueous Fraction with invivo Response Absorbance, Ranking of in vivo Sample ID mAU/g, 476 nmlutein response AET-C 57.6 1 (lowest response) AET-1 62.7 4 AET-2 61.3 3AET-3 59.4 2 AET-4 64.9 5 (highest response)

As may be seen from the table, the absorbance values correlate with thein vivo results, in that each of the nutritional compositions comprisingan activated MDG premix (AET-2 and AET-4) or an MDG-protected premix(AET-1) have higher lutein absorbance values than do the controlnutritional composition (AET-C) or the nutritional composition that doesnot comprise an activated premix (AET-3). More lutein is present in theaqueous fraction of the nutritional compositions comprising an activatedMDG premix or an MDG-protected premix. Thus, the relativebioavailability of lutein, as demonstrated by in vivo testing, wasaccurately predicted by in vitro testing using this method.

Example 56

In this Example, the effect on the bioavailability of DHA in nutritionalcompositions containing an activated MDG premix or an MDG-protectedpremix is evaluated and compared to the bioavailability of DHA that isin nutritional compositions without the activated MDG premix or theMDG-protected premix.

A similar protocol for animal infusion and lymph collection as describedin Example 1 was followed, including the infusion of the samenutritional compositions (e.g., compositions AET-C, AET-1, AET-2, AET-3and AET-4). The lymph lipids were extracted and analyzed for DHAconcentration using high-performance liquid chromatography. FIG. 22shows the change in lymphatic output of DHA over time. There was asignificant increase in lymph levels of DHA from the initial (fasting)levels to the DHA levels in the 6 hrs following infusion for thenutritional compositions containing an activated premix (AET-2) or anMDG-protected premix (AET-1). In comparison, the control nutritionalcomposition (AET-C) and the nutritional composition that does notcomprise an activated premix (AET-3) showed only a modest increase inthe lymph levels of DHA over time.

Example 57

A study was conducted to compare a commercially representativenutritional powder with and without MDG. Nutritional powder batches(control and MDG) were reconstituted (152 g/L), centrifuged(31,000×g/20° C./4 h), and the MDG concentration in the aqueous fractionof samples was estimated by HPLC determination of monolinolein andmonoolein, and represented as mg of MDG per kg of aqueous fraction.

TABLE 39 MDG presence in the Aqueous Fraction after NutritionalComposition Centrifugation Aug. 15, Sep. 19, Nov. 5, Feb. 15, Batch 20132013 2013 2014 AET-C <70 <40 <20 NT (no MDG) (not tested) AET-1 144 120NT NT AET-2 NT 150 NT NT AET-3 NT 90 118 NT AET-4 178 140 115 NT AET4-5NT NT 127 NT A455 NT NT NT <10   (no MDG) A456 NT NT NT 30 A457 NT NT NT40 A458 NT NT NT 40

A measurable presence of MDG in the aqueous fractions of all MDG sampleswas detected, and an absence of MDG in the aqueous fraction of allcontrol batches, was verified by LC/UV analysis. The significance of thedata is that (a) MDG is present in the Aqueous Fraction (despite itsvery low water solubility), and that (b) the MDG concentration isgenerally correlated with in vivo lutein availability.

Example 58

A study was conducted to investigate the distribution of lutein and MDGwithin the nutritional compositions. Sample were prepared usinghomogenization at 150/20 bar, and then centrifuged at 100,000×g for 1hour. The different layers (e.g., cream, aqueous, and pellet) were thenanalyzed for the presence of lutein and MDG.

The results demonstrate that nutritional compositions comprising MDGprovided MDG and lutein within the aqueous phase, wherein thesemolecules have very limited water solubility. Specifically, it was foundthat lutein and MDG were present in the aqueous phase, followingcentrifugation, at approximately 4 wt % and approximately 8 wt % of thenutritional composition respectively. This suggests that the luteinfound in the aqueous phase is associated with water-soluble complexes.It should be noted that the pellet contained approximately 1% of thelutein, suggesting that lutein could be associated with a sedimentableassembly as well.

Example 59

A study was conducted to investigate the dispersibility/solubility ofMDG (mono- and diglycerides) in various aqueous preparations asevaluated by HPLC. The MDG was added to water at 95° C., RT, and thenthe samples were subjected to high speed centrifugation at 20° C., andare reported in the table below.

TABLE 40 MDG Alone has Limited Solubility in the Aqueous FractionSoluble MDG, Soluble MDG, as % MDG Preparation as mg per kg of total MDG1.00% (w/w) MDG 1980 19.8% suspension in 95° C. water Room temperature~32 ~0.32% filtrate of 1.00% (w/w) MDG suspension Centrifugation ~16~0.16% supernatant of room temperature filtrate

The MDG present following centrifugation was very limited, and is instark contrast to the MDG present within the aqueous phase followingcentrifugation of the nutritional composition. This suggests that thesolubility of MDG, along with the lipophilic compound, is increased innutritional compositions comprising MDG, at least one lipophiliccompound, and at least one hydrophobic protein.

Example 60

A study was conducted to further investigate the distribution of luteinand MDG within the nutritional composition. Samples includedMDG/lutein/water; MDG/lutein/NFDM/water; MDG/lutein/WPC/water; andMDG/water.

The premixes were provided as follows: in samples that contained lutein,lutein was heated for 2 h at 50° C. with shaking. In samples thatcontained MDG (GMO-40) and lutein, lutein was added to MDG and heated to50-60° C. and mixed for 10 minutes. In samples that contained protein,non-fat dried milk was added to water (50-60° C.) with agitation andallowed to hydrate for approximately 10 min.

The samples were prepared as follows: for solution 1, water was heatedto 50-60° C., an lutein in GMO-40 blend and turrax (30 sec 10000 rpm)was added; for solution 2, NFDM was placed in water solution heated to50-60° C., and lutein was added in GMO-40 blend and turrax (30 sec 10000rpm); for solution 3, water was heated to 50-60° C., and GMO-40 blendand turrax (30 sec 10000 rpm) was added. The solutions are listed in thetable below as X.1, wherein 1 is the solution.

TABLE 41 Batch Details for Example 60 sample MDG + MDG + MDG + lutein +lutein + lutein NFDM WPC MDG 3.1 3.2 3.2-1 3.2-2 3.3 lutein 0.06950.0601 0.0601 0.0601 0 GMO-40 9.18 7.94 7.94 7.94 9.18 dried milk 0107.98 107.98 0 0 (nonfat dried milk, NFDM) Whey protein 0 0 0 107.98 0concentrate 80 (WPC) water 790.75 684.02 684.02 684.02 790.82 TOTAL [g]800 800 800 800 800

Samples were formulated as detailed above, and then characterized byultrafiltration with various membranes, as well as ultracentrifuged atvarious times and speed (e.g., 100,000×g, 4,500×g, and 1,000×g). Thedetails of the different centrifugation/filtration method are listedbelow.

Speed of centrifugation: 1000×g, Centrifugation time: 15 min,Centrifugation temperature: 20° C., Centrifuge tubes: Cellstar tube 50ml from Greiner bio-one, Tube diameter: 23 mm, Amount of product intube: 45 ml, Centrifuge: type 5810 R from Eppendorf, aqueous phaseisolation by syringe through the cream layer.

Speed of centrifugation: 4500×g, Centrifugation time: 15 min,Centrifugation temperature: 20° C., Centrifuge tubes: Cellstar tube 50ml from Greiner bio-one, Tube diameter: 23 mm, Amount of product intube: 45 ml, Centrifuge: type 5810 R from Eppendorf, aqueous phase bysyringe through the cream layer.

Speed of centrifugation: 100,000×g, Centrifugation time: 1 h,Centrifugation temperature: 20° C., Centrifuge tubes: Ultra ClearCentrifuge tubes from Beckmann Coulter, Tube size: 14×89 mm, Amount ofproduct in tube: 11.3 g, Centrifuge: type L-90K from Beckmann Coulter,aqueous phase isolation by freezing the tube with content in liquidnitrogen and cutting the part with the serum phase including the tubewall followed by thawing the content of this part of the tube.

Membrane: Centrifugal filter units from Amicon Ultra; 100 kDa and 10kDa, Centrifugation speed: 4000 g, Centrifugation time: 30 min,Centrifugation temperature: 20° C., Centrifuge tubes: Cellstar tube 50ml from Greiner bio-one, Tube diameter: 23 mm, Amount of product intube: 45 ml, Centrifuge: type 5810 R from Eppendorf, Serum isolation byremoving the filter form the tube and decanting the filtrate.

Centrifugation speed (Lumifuge): 1000 g, Centrifugation time: 255 min,Light factor: 1, Centrifugation temperature: 20° C., Lumifuge: typeCentrifuge lumiview 110 from LUM Gmbh.

Following centrifugation and/or ultrafiltration, samples were analyzedfor the presence of lutein, and MDG within the different phases.

Lutein analysis as done by RP-HPLC-DAD, and MDG analysis was done byRP-HPLC-UV-ELSD. In MDG analysis, samples were prepared as follows:samples were weighed at 0.15 g in a 20 mL tell bottle; the solution wasstirred on a magnetic stirrer in the fume hood; 15.0 mL or 13.5 mL, 80%acetone+20% chloroform were added with a measuring cylinder.

Stir the solution for 10 minutes on a magnetic stirrer in a water bathat ±50° C. (warm water from the tap); the solution was sonicated for 10minutes at room temperature; 1.5 mL of sample was centrifuged in a 1.5mL centrifuge tube, 10 min at 20.000×g; 20 uL of the supernatant wasinjected on the HPLC column. UV spectroscopy of the samples wasperformed with wavelength scans (200-900 nm at 5 nm intervals) at 20° C.using a Cary 4000 UV-Vis Spectrophotometer (Varian BV). All measurementswere carried out in quartz cuvettes with a 1 mm path length.

Qualitatively, samples without protein resulted in more appreciableamounts of precipitated lutein along the side of the sample flasks. Thissuggests that there is an optimal interaction between lutein,hydrophobic protein and MDG to achieve the water soluble assembly.

In the ultrafiltration analysis, no lutein or MDG passed through the 100kD filter, this was across all samples (FIG. 23).

The results also demonstrated that the presence of protein resulted inmore lutein and MDG in the aqueous phase. Furthermore, when comparingWPC vs. NFDM, WPC resulted in both more lutein and MDG in the aqueousphase. There may be different particle characteristics (e.g., size) offor the WPC batch, that results in greater distribution of lutein andMDG in the aqueous phase, relative to NFDM.

Centrifugation at 100,000×g: lutein was only found in the aqueous phaseof samples containing protein; negligible lutein was found in aqueousphases of samples containing only lutein and MDG.

Centrifugation at 4,500×g: similar amounts of lutein were found in theaqueous phase of samples containing NFDM and WPC. More MDG was found inthe aqueous phase of samples containing WPC than NFDM. Additionally,concentrations of lutein and MDG in the aqueous were considerably higherthan in those of protein-free systems centrifuged at 1,000×g.

Centrifugation at 1,000×g: resulted in considerably higher amounts oflutein in the aqueous phase of samples with NFDM, than without.Specifically, ˜20% lutein was found in the aqueous phase at thiscentrifugation speed. Overall, the data suggest that there is an optimalinteraction between lutein, hydrophobic protein and MDG to achieve thewater soluble assembly.

TABLE 42 Centrifugation/Ultrafiltration Analysis of Variable MDG Batches3.1 3.2 3.2-1 3.2-2 3.3 sample Lutein MDG Lutein MDG Lutein MDG LuteinMDG Lutein MDG Ultrafiltration n.a. n.a. n.a. n.a. n.d. n.d. n.d. n.d.n.a. n.a. Centrifugation 0.1 0.047 3.3 3.572 n.d. n.d. n.d. n.d. n.a0.234 method A (1000 g) Centrifugation n.d. n.d. n.d. n.d.  1.6 1.967 1.6 3.01 n.d. n.d. method B (4500 g) Centrifugation n.a. n.a 0.2 1.59 n.d. n.d. ad. n.a. n.a method C (100,000 g) Overall 13.1 7.33  15.4 8.44sample

Example 61

A study was conducted to investigate the effect of shelf-life on MGDbatches. Different MDG bathes included, 18 month old batch, 24 month oldbatch (AET-1), and the control. The different MDG batches were examinedin the lymphatic model as described in Example 20.

The results demonstrate that the older batch of bounty performed atsimilar levels as the newer batch in the bioavailability of lutein (FIG.24). This suggests that the assembly of the nutritional composition isstable for at least 24 months.

1. A nutritional composition having at least one protein, at least onefat, and at least one lipophilic compound, the composition comprising:an activated assembly comprising at least one hydrophobic protein, atleast one combination of monoglycerides and diglycerides (“MDG”) and atleast one lipophilic compound, wherein at least 1% of the total MDG inthe nutritional composition remains in the aqueous phase aftercentrifugation at 100,000×g for 1 hour at 20° C.
 2. The nutritionalcomposition of claim 1, wherein the assembly is (i) water soluble; and(ii) has a size of 100 kD to 1000 kD.
 3. The nutritional composition ofclaim 1, wherein MDG increases the concentration of protein of theassembly by at least 2% compared to a nutritional composition withoutMDG.
 4. The nutritional composition of claim 1, wherein MDG increasesthe bound lipophilic compound of the assembly by at least 2% compared toa nutritional composition without MDG.
 5. (canceled)
 6. The nutritionalcomposition of claim 1, wherein the assembly is still water solubleafter centrifugation at about 31,000×g for at least 1 hour at 20° C.7-8. (canceled)
 9. The nutritional composition of claim 1, wherein theassembly is stable for at least 12 months at 23° C. 10-11. (canceled)12. The nutritional composition of claim 1, wherein the hydrophobicprotein is β-lactoglobulin, bovine lactoferrin, γ₂-casein, β-casein,α-lactalbumin, or combinations thereof.
 13. (canceled)
 14. Thenutritional composition of claim 1, wherein the lipophilic compound is acarotenoid, a lipid soluble vitamin, a lipophilic antioxidant orcombinations thereof.
 15. (canceled)
 16. The nutritional composition ofclaim 1, wherein the fat is MDG, coconut oil, fractionated coconut oil,soy oil, corn oil, olive oil, safflower oil, high oleic coconut oil,fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil,high oleic safflower oil, sunflower oil, high oleic sunflower oil, palmoil, palm kernel oil, palm olein, canola oil, marine oils, cottonseedoil, flax seed oil, hemp seed oil, peanut oil, borage oil, algal oils,fungal oils, MCT oil (medium chain triglycerides) and combinationsthereof.
 17. The nutritional composition of claim 1 comprising about 145mg/kg to about 890 mg/kg of MDG based on total weight of thecomposition.
 18. (canceled)
 19. The nutritional composition of claim 1comprising about 1.0 μg/kg to about 7.0 g/kg of a lipophilic compoundbased on total weight of the composition. 20-21. (canceled)
 22. Thenutritional composition of claim 1, wherein the ratio of MDG tolipophilic compound comprises a ratio of about 12000:1 to about 1:1(w/w).
 23. (canceled)
 24. The nutritional composition of claim 1comprising about 1.0 g/L to about 100 g/L of hydrophobic protein basedon total weight of the composition.
 25. The nutritional composition ofclaim 1 further comprising choline.
 26. (canceled)
 27. The nutritionalcomposition of claim 1 further comprising lecithin.
 28. (canceled) 29.The nutritional composition of claim 1 further comprisingdocosahexaenoic acid (“DHA”).
 30. (canceled)
 31. The nutritionalcomposition of claim 1 further comprising high palmitic acid vegetableoil. 32-33. (canceled)
 34. The nutritional composition of claim 1,wherein the nutritional composition comprises a carbohydrate. 35.(canceled)
 36. A nutritional composition comprising at least one proteinand vitamin D, having improved bioavailability of vitamin D, wherein thenutritional composition comprises: an assembly comprising a combinationof monoglycerides and diglycerides (“MDG”) and vitamin D, wherein thevitamin D has improved bioavailability when administered to a subject.37-40. (canceled)
 41. A nutritional composition having at least oneprotein, at least one fat, and at least one lipophilic compound, thecomposition comprising: an activated assembly comprising at least onehydrophobic protein, at least one combination of monoglycerides anddiglycerides (“MDG”) and at least one lipophilic compound, wherein atleast 15% of the total lipophilic compound in the nutritionalcomposition remains in the aqueous phase after centrifugation at 1,000×gfor 1 hour at 20° C.